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PATHOLOGY
HANDBOOK
Capital Pathology ABN 49 452 500 422
2 Makin Place, Deakin, ACT, 2600, Australia
All rights reserved. No part of this manual may be reproduced,
stored in a retrieval system, or transmitted in any form or any
means without Capital Pathology’s written consent.
This Handbook has been prepared by Capital Pathology and every
care has been taken in its compilation. Capital Pathology shall not
be liable for any error or omission contained herein or arising from
usage of material contained in this Handbook.
This Handbook remains the property of Capital Pathology.
Published–2012
Edited by Christina Vett-Joice
Printed by Buckner Printing Company
i
Table of Contents
Acknowledgementsiii
Forewordiv
How to use this handbook
v
General information
Pathologistsvi
Overview
xvi
Services
xvii
Contact Us
xviii
Doctors Service Centre
xx
Collection Centres
xxi
Specimen Collection
xxiii
Courier Services
xxiv
Requesting Pathology Tests
xxv
Laboratories
xxvii
Results
xxviii
Reports
xxix
xxxi
Billing Policy
Education
xxxiv
Publications and website
xxxv
Other Services
xxxvi
Corporate services
xxxvii
A–Z Listings
ii
Acknowledgements
This is the fifth edition of the Capital Pathology handbook, and the production of each new
edition is a huge task. Also, each edition builds on the contributions that others have made
to earlier editions so that the current handbook is a testament to the hard work of many
teams of people over many years.
There have been champions of the fifth edition handbook within Capital Pathology, just as
there were champions of its predecessors.
Dr Paul Whiting, Director of Clinical Pathology has again been passionate about the
urgency of keeping the handbook relevant and current for our referring doctors and both
myself and Dr Jane Twin, Director of Cytology have also contributed extensively to our
relevant sections.
Christina Vett–Joice (CV) has once again been the driving force behind the fifth edition.
She has been cheerfully assisted by the scientific and administration staff of
Capital Pathology, many of whom have helped produce parts of this handbook in their
own precious non-work time.
Many people throughout Sonic Healthcare have been incredibly generous in their
contributions to both this and earlier editions. They have shared their knowledge,
experience and advice with us and we thank you. It is wonderful to have a large team of
world experts at our disposal at Capital Pathology and we will continue to test the limits of
your kindness and availability!
I would also like to thank the companies who have sponsored the fifth edition. Without your
generosity, we would not have been able to produce a reference work of such quality and
scope, and we appreciate your faith in our handbook and your support to us.
Finally I would like to sincerely thank those who have fundamentally made this handbook
possible through their ongoing support and loyalty to Capital Pathology –
the doctors, nurses and patients who daily trust us with the privilege of being part of their
health care team. I assure you that we take our responsibilities to you very seriously. In fact
“We Take it Personally”.
Dr Ian Clark
M.B., B.S., F.R.C.P.A., M.I.A.C., F.A.S.C.P., F.A.I.C.D.
Chief Executive Officer
2012
iii
Foreword
We are constantly reminded that we are in the midst of an explosion of knowledge and
information. This is nowhere typified better than in the ever-changing field of pathology.
This is the fifth edition of the handbook provided by Capital Pathology to medical
practitioners, nurses, practice managers, receptionists, hospital staff and others who use
our practice. The handbook provides a concise guide to the range of services provided by
Capital Pathology, as well as assisting to answer some of the clinical questions we are so
often asked.
Capital Pathology has a long history, over 40 years, of providing pathology services to
the ACT and surrounding districts. We are very aware of the privilege and responsibility
associated with being an integral part of the medical community in our region, and this
can be summarised in our motto “We take it personally”.
For us in the potentially rarefied atmosphere of an efficient modern pathology laboratory,
this means that we constantly remind ourselves that associated with every specimen,
every report, every piece of anonymous data, every phone query, there is a person who
rightfully expects us to treat them “personally”.
This also means that our supporting doctors and their teams can rely on each of us at
Capital Pathology, irrespective of our role, to behave as part of a medical practice where
we are each known by you, and are personally committed to ensuring the best possible
service for you and your practice.
I hope that this Handbook will help us improve our service to you. I also hope it will assist
you and your staff to achieve the ultimate aim of us all: to improve the health of our
patients and our community.
Dr Ian Clark
M.B., B.S., F.R.C.P.A., M.I.A.C., F.A.S.C.P., F.A.I.C.D.
Chief Executive Officer
2012
iv
How to use this handbook
All listings are in alphabetical order, whether analyte, test name, disease, clinical topic,
procedure, drug name, specimen type or pathogen.
Reference Ranges
For some common analytes the reference ranges are similar throughout Australia
(e.g. sodium, potassium). For hormones and other tests where methodology can vary
from one laboratory to another, the reference ranges can differ quite markedly. Even in the
same laboratory, ranges may change from time to time as new methods are introduced. In
this handbook you are provided with common reference ranges.
For other tests, please consult the reference ranges provided with all reports.
Please note reference ranges are correct at time of printing.
A reference range can be defined as a range derived from measuring an analyte in a
reference population using a reference method. The reference interval then is usually
taken to include the mean +/–2 SD i.e. 95th percentile. This implies that 5% of the normal
population will fall outside these limits.
In clinical practice, a patient’s results can be examined in different ways, either by
comparison to a reference interval, by comparison to their own known previous values, or
by consideration of desirable goals, e.g. aiming for a cholesterol of X since this has been
shown to have benefit in terms of mortality / morbidity from cardiovascular disease. Some
useful ranges are not reference ranges at all, but rather therapeutic ranges, e.g. warfarin
control by INR.
Units
S.I. units are used throughout.
Notes on Interpretation
Comments in this handbook are intended as a guide only. Any additional queries may be
directed to the appropriate pathologist.
Brief clinical details help immeasurably with interpretation,
e.g. routine, wt–loss 1/12, pale and tired 3/12, on T4, on phenytoin, ? hepatitis.
v
Pathologists
In my opinion, pathologists are undoubtedly the central focus of a successful pathology
practice.
At Capital Pathology, specialist pathologists are not only responsible for management
decisions, but are intimately involved in all clinical and ethical matters. Our pathologists
pride themselves on their close clinical relationship with our referring doctors, and they
recognise the importance of providing a comprehensive consultative and educational
service. This is exemplified on a day-to-day basis by an expanding range of interpretive
comments on reports. In order to optimise patient care, we understand the importance of
discussion with clinicians through phone calls as well as providing advice and feedback
at personal visits. Capital Pathology’s specialist pathologists have a long history of
commitment to continuing education for family doctors and specialists through educational
seminars and regular newsletters.
Even more importantly, the continual guidance provided by our pathologists determines
the quality and integrity of our whole service. Although our practice obviously must remain
commercially viable, the medical influence provided by our pathologists ensures that
purely commercial considerations never take precedence over ethical and professional
standards.
Dr Ian Clark
M.B., B.S., F.R.C.P.A., M.I.A.C., F.A.S.C.P., F.A.I.C.D.
Chief Executive Officer
vi
Dr Ian Clark
M.B., B.S. (Hons), F.R.C.P.A., M.I.A.C., F.A.S.C.P., F.A.I.C.D.
Chief Executive Officer
Director of Histopathology
Ian graduated with first class honours from Sydney University
in 1984. He was full–time Senior Tutor in the Pathology
Department of Sydney University before commencing his
Registrarship in Histopathology at the Royal North Shore
Hospital of Sydney, where he also developed expertise in
fine needle aspiration cytology. He completed his Fellowship
examinations in 1990, after which he joined the Douglass
Laboratories practice in Sydney as a full–time specialist
pathologist.
In 1996 Ian was appointed Deputy Director of Histopathology
of the newly merged Douglass Laboratories and Hanly Moir
Pathology. He joined Capital Pathology in July 1998 as
Director of Histopathology, was appointed Medical Director in
2001 and became CEO in 2007. In 2005 Ian was appointed
Senior Clinical Lecturer at the ANU Medical School.
Ian is a Histopathologist and Cytopathologist with keen
special interests in dermatological, gastrointestinal,
gynaecological and genito–urinary pathology. He also
oversees the application of flow cytometry to cytological
and histological specimens.
Ian is an active member of many medical associations and
societies, including the Australian Society for Colposcopy
and Cervical Pathology, the Australasian Society for Breast
Disease and the Australian Dermatopathology Society. He
holds Membership of the International Academy of Cytology
and is a Foreign Fellow of the American Society of Clinical
Pathologists.
He has participated in a number of research projects, and
has spoken at many conferences, including the International
Academy of Pathology. Ian was President of the Australian
Association of Pathology Practices from 2007 to 2009 and is
the Honorary Secretary of the Royal College of Pathologists
of Australia. He is also the Pathology craft representative
on the ACT AMA.
The fundamental importance of medical leadership underpins
Ian’s role as Chief Executive Officer, and he is committed to
maintaining the high quality and ethical standards for which
the practice is renown.
If you have any enquiries for Ian, please do not hesitate to
contact him on 02 6285 9800.
vii
Dr Jane Twin
M.B., B.S., F.R.C.P.A., F.I.A.C., Dip. Cytopath.
Director of Cytopathology
Jane is a graduate of the University of Tasmania and
undertook her Pathology training at Royal Hobart Hospital.
Following attainment of the Fellowship of the Royal College
of Pathologists of Australasia in 1983, Jane worked as a
researcher and lecturer at the University of Tasmania with
an appointment to the Royal Hobart Hospital as a provider
of Cytology Services. After training with Dr Svante Orell in
Adelaide, Jane moved to Hobart Pathology in 1989 to set up
a Fine Needle Aspiration Service. This service grew rapidly
and was augmented by provision of FNA and Histopathology
to BreastScreen Tasmania. Other involvement in
BreastScreen Tasmania included her appointment as
Tasmania’s representative on the National Pathology Q–
group and a long term role as Pathology adviser and auditor.
Jane was also involved in the setting up of the Tasmanian
Cervical Cytology Register and continued on the Technical
Working Party until leaving Tasmania in December 2001.
Other achievements include attainment of further
qualifications in Cytopathology by examination, including
Fellowship of the International Academy of Cytology (1992)
and the Royal College of Pathologists of Australasia Diploma
of Cytopathology (1997). Jane has been involved with the
Australian Society of Cytology at a National Level since 1991
as a Councillor and on the Board of Examiners, and has a
strong interest and experience in Gynaecological Pathology.
Jane is a member of the advisory committee for the Cytology
Quality Assurance Program and is also a member of the
RCPA cytology discipline advisory committee. She is also
a member of the ACT Papsmear Register management
committee. At Capital Pathology, Jane continues her
interests in cytology and the histological aspects of
gynaecological and breast pathology while contributing
generally to the histology department.
If you have any enquiries for Jane, please do not hesitate to
contact her on 02 6285 9867.
viii
Dr Paul Whiting
M.B., B.S., F.R.C.P.A., M.A.S.M.
Director of Clinical Pathology
Paul studied for his medical degree at the University of
Melbourne, graduating in 1987. After residency and a
year in General Practice, Paul commenced his pathology
training at Geelong Hospital. He moved to Canberra in 1996
where he completed his general pathology training. He was
admitted as a Fellow of the Royal College of Pathologists
of Australasia ( RCPA ) before joining Capital Pathology as
the Director of Clinical Pathology in 1998. In this role, he
supervises the haematology, biochemistry, immunology and
microbiology laboratories, and is responsible for ensuring
that the highest quality standards are always met. Paul
encourages consultation on the wide variety of problems
in clinical medicine, and is always available to discuss
pathology results.
Paul is involved with many professional associations and is
on several hospital advisory committees. He is a member
of the Haematology Society of Australia, the Australian
Association of Clinical Biochemists, the Australian Society
for Microbiology and is also a member of the American
Society of Clinical Pathologists. Paul is active with many
quality assurance interests, and acts as a referee in the
RCPA external quality assurance programs. In Canberra
he maintains a continuing interest in medical education and
regularly presents to local specialist and general practitioner
audiences.
If you have any enquiries for Paul, please do not hesitate to
contact him on 02 6285 9895.
ix
Dr John Docker
M.B., B.S., D.C.P. (London),.D.R.C. Path (UK), F.R.C.P.A.
Specialist Pathologist
John graduated from Sydney University in 1969 and did
his early training in general pathology at the Charing Cross
Hospital in London, where he was subsequently appointed
Senior Registrar and Lecturer at the Charing Cross Medical
School. John returned to Sydney with his young family
in 1975 and after a year at St. Vincent’s Hospital and the
Prince Henry / Prince of Wales Hospitals, he qualified for his
Fellowship and returned to his boyhood town of Goulburn in
1978. Since then he has practised in Goulburn as a General
Pathologist with a strong emphasis on histopathology. John
joined Capital Pathology on a part-time basis in June 1999.
If you have any enquiries for John, please do not hesitate
to contact him on 02 6285 9867.
x
Dr Juli Ferguson
M.D., Ph.D., F.C.A.P.
Specialist Histopathologist
Juli graduated with a BS cum laude from the College of
St. Mary, Omaha, Nebraska, U.S. in 1971. She entered
graduate school completing her MS (1974) and Ph.D. (1977)
in the field of Biochemistry at the University of Nebraska.
She then entered medical school, completing her M.D.
program in 1981 with acceptance into the Alpha Omega
Alpha medical honour society. After her Registrarship at
the University of Iowa Hospitals and Clinics, she joined
Physicians Associates in Grand Island, Nebraska in
1985. She became a fellow of the College of American
Pathologists in 1989.
Juli has been with Capital Pathology since immigration
to Australia in 1989. Juli’s special interests are urologic
pathology and dermatopathology. She is a member of
the American Society of Clinical Pathologists, College of
American Dermatopathology Society and International
Academy of Science.
If you have any enquiries for Juli, please do not hesitate to
contact her on 02 6285 9867.
xi
Dr Peter Harper
M.B., Ch.B.(N.Z.), F.R.C.P.A.
Specialist Histopathologist
Dr Peter Harper graduated from the University of
New Zealand (Otago) and his residency was undertaken
in large and small teaching Hospitals and a year in remote
General Practice before commencing his Pathology career. At the beginning of 1968 he migrated to Australia pursuing
Pathology under Dr Baird in Biochemistry and Haematology
at the Royal Melbourne Hospital. At various times he
has been tutor in Pathology at Melbourne and Monash
Universities. He then entered Private Pathology Practice selling his own
Practice in 1987 and joining Gribbles Pathology as Head of
Histopathology. He undertook a stint in country NSW and
Victorian public and Private Practice and was a Coronial
Pathologist in NSW and Victoria.
He again entered private practice in 1992 with Macquarie
Pathology and Mayne Health and in 2005 joined SDS prior
to joining Capital Pathology in Canberra in 2008.
Peter has a keen interest in skin pathology, and welcomes
discussion with referring doctors. He can be contacted on
02 6285 9867.
xii
Dr Tracey Lu
M.B., F.R.C.P.A., B.M.L.S
Specialist Histopathologist and Cytopathologist
Tracey has dual degrees in both Medicine and Medical
Laboratory Science. She obtained her primary medical
degree from Fujian Medical University China in 1986 and
worked there when she graduated as a full-time assistant
lecturer, researcher and clinical anatomical pathologist
before immigrating to New Zealand.
After settling in NZ, Tracey gained a Bachelor degree in
Medical Laboratory Science from Auckland University
of Technology and worked as a Scientist at Labplus,
Auckland Hospital whilst she passed the New Zealand
Medical Registration Exam (NZREX). She then worked
as a House Surgeon at a New Zealand teaching hospital
before commencing her Pathology Registraship in 2004.
Tracey trained in Anatomical Pathology in Auckland and
Tauranga NZ for a total of five years. She worked in rotations
between three public hospitals and one private laboratory
and gained experience in a wide variety of subspecialties,
including cytopathology, breast, gynaecology, urology, GI,
cardiothoracic, neuropathology, head and neck, bone and
soft tissue, paediatric, forensic, and lymphoma pathology.
Tracey has joined Capital Pathology as a specialist
Histopathologist and Cytopathologist after she was awarded
her Fellowship from The Royal College of Pathologists of
Australasia.
If you have any enquiries for Tracey, please do not hesitate
to contact her on 02 6285 9867.
xiii
Dr Sumi Ranjit
MBBS, DCH, MRCP (Paediatrics), FRCPA (Haematology),
FRCPA (Anat Path)
Specialist Haematologist and Anatomical Pathologist
Sumi brings with her a rich and diverse medical background
stretching over a period of more than 20 years.
After obtaining her primary medical qualification and training
in Paediatrics from Madras University in India, she moved
to the UK to undertake training in Paediatric Haematology
at the Royal Manchester Children’s Hospital and St James’s
University Hospital in Leeds. Her Haematology skills were
further honed while working as Registrar and Research
Fellow at Prince of Wales Hospital in Sydney (1999–2002).
After obtaining her specialist qualifications in Haematology
(FRCPA) in 2002, Sumi worked as a specialist Haematologist
in Canberra and overseas. However, in order to fulfil a long
felt desire to gain more in depth knowledge, Sumi undertook
further training in Anatomical Pathology at The Canberra
Hospital and at Capital Pathology, passing her final
examinations and obtaining her Fellowship in Anatomical
Pathology in 2010.
Sumi co-edited a Handbook of Common Clinical Emergencies
while still a medical student in India. More recently, she has
made several presentations at national and international
meetings, including the American Society of Haematology
and the Australian Society of Cytopathology. She has also
held the post of Associate Lecturer at the Australian National
University Medical School where she was involved in
teaching medical students.
Sumi and her family enjoy living in Canberra and she is
passionate about the environment. Her other interests
include reading, travelling, cooking and natural history.
If you have any enquiries for Sumi, please do not hesitate
to contact her on 02 6285 9867.
xiv
Dr Melissa Robbie
M.B.B.S., F.R.C.P.A.
Specialist Histopathologist and Cytopathologist
After graduating from medical school at the University
of Melbourne in 1983, Melissa began her post-graduate
training at Queen Victoria Hospital (later Monash Medical
Centre), where she entered the Pathology training program.
This was followed by further registrar positions at Monash
Medical Centre and Dandenong District Hospital. She
received her fellowship from the Australian Royal College
of Pathologists in 1991 and had her first consultant
appointment in the General Pathology Service at Ballarat
Base Hospital. Melissa has a keen interest in teaching and
subsequently spent three years teaching 2nd and 4th year
medical students at Monash Medical University Medical
School, as well as consulting in Anatomical Pathology
at Dandenong Hospital and Eastern Suburbs Pathology
Services.
From 1995 until 2000, Melissa was Senior Pathologist /
Deputy Director of Pathology at Mercy Hospital for Women
& Werribee Mercy Hospital. From 2000 to early 2005, she
worked at St Vincent’s Hospital, Melbourne. In addition,
Melissa continues to hold an honorary appointment at
Peter MacCallum Cancer Institute, where for 9 years she
was central reviewer for Gynaecologic Pathology and now
has continuing research collaborations investigating the
mechanisms of breast and ovarian cancer.
Melissa has special interests in Gynaecological & Obstetric
Pathology, Cytopathology, and Gastro-intestinal pathology.
She is a member of the Australian Cancer Network’s
committee on ovarian cancer, which recently produced the
first national guidelines on ovarian cancer management for
doctors and patients. Melissa also contributed a chapter
to an international collaborative work on Controversies in
Gynecologic Cancer, edited by David Gershenson et al
(Churchill Livingstone 2004) and is now a member of the
Editorial board for the journal Gynecologic Oncology.
If you have any enquiries for Melissa, please do not hesitate
to contact her on 02 6285 9867.
xv
Overview
Capital Pathology is a fully NATA accredited Category GX medical testing laboratory
that provides quality pathology services to general practitioners, medical specialists,
nursing homes, private hospitals and the community in the ACT, South Coast,
Snowy Mountains and Goulburn regions.
We are proud to have provided services to the ACT and adjoining regions for over
40 years.
Our purpose built, state of the art main laboratory is situated in Deakin, right in the
heart of Canberra. We also have regional laboratories conveniently located in both
Bega and Goulburn. Our practice currently employs in excess of 300 local staff.
Our practice encompasses all disciplines of pathology. Our specialist pathologists,
scientists and technical staff are available for consultation and general enquiries.
Capital Pathology is a division of Sonic Healthcare Limited, an Australian owned,
publicly listed company.
Our Mission
We help doctors help patients by providing specialist pathology services.
Our Values
Commit to Service Excellence
To willingly serve all those with whom we deal with unsurpassed excellence.
Treat Each Other with Respect and Honesty
To grow a workplace where trust, team spirit and equity are an integral part of
everything we do.
Demonstrate Responsibility and Accountability
To set an example, to take ownership of each situation to the best of our ability
and to seek help when needed.
Be Enthusiastic about Continuous Improvement
To never be complacent, to recognise limitations and opportunities for ourselves
and processes and to learn through these.
Maintain Confidentiality
To keep all information pertaining to patients as well as professional and commercial
issues, in strict confidence.
xvi
Services
Mission Statement
‘We help doctors help patients by providing specialist pathology services’
We provide a comprehensive range of pathology testing for doctors and their patients
serving:
•
•
•
•
•
General Practitioners
Specialists
Private Hospitals
Nursing Homes, Aged Care, Supported Care
Day Surgeries.
Our laboratory offers:
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Main laboratory open 24 hours, 7 days a week
Specialised Pathologists on-site
Doctors Service Centre dedicated to providing results and assisting with enquiries
A comprehensive range of routine and emergency pathology testing
Fine needle aspiration service available on-site
Regional laboratories in Bega and Goulburn
Rapid turn around of results
Innovative report formats
Personalised options in reporting
Statistical analysis of various reports available
Conveniently located collection centres in the local and surrounding districts open
extended hours and offering appointments
Home visits available if required
Extensive courier network
Electronic access and downloading of results
Publications and ongoing education for referring practitioners and their staff.
We also offer a broad range of services for corporate and industrial customers including:
•
•
•
•
Occupational and environmental testing
Insurance and corporate testing
Clinical trials
Food and water testing.
Our staff are always available to support practitioners in their clinical practice.
Our Pathologists encourage doctors to contact them for information or advice at any time.
Our Client Services Department is available to assist all our customers with enquiries,
issues and information.
xvii
Contact Us
2 Makin Place, Deakin, ACT, 2600
The laboratory provides services 24 hours a day, 7 days a week.
Doctors Service Centre (DSC)
Results and General Enquiries......................................................................02 6285 9803
Front Reception Switchboard.........................................................................02 6285 9800
Accounts ................................................................................................... 02 6285 9888
Chief Executive Officer.............................................................................. 02 6285 9836
Client Services........................................................................ 02 6285 9805/9802/9801
Collection................................................................................................... 02 6285 9881
Couriers..................................................................................................... 02 6285 9877
Cytology.................................................................................................... 02 6285 9868
Histology.................................................................................................... 02 6285 9867
Home Visits:
Northside .................................................................................................. 02 6251 5121
Southside ................................................................................................. 02 6281 7277
Central...................................................................................................... 02 6285 9885
xviii
IT Services................................................................................................. 02 6285 9860
Main Laboratory ........................................................................................02 6285 9811
Microbiology ............................................................................................. 02 6285 9846
Pathologists
CEO/Medical Director/Director of Histopathology.......Dr Ian Clark .......... 02 6285 9800
Director of Clinical Pathology......................................Dr Paul Whiting..... 02 6285 9895
Director of Cytopathology............................................Dr Jane Twin......... 02 6285 9867
Stores ...................................................................................................... 02 6285 9813
Fax
Administration............................................................................................ 02 6281 1941
Doctors Service Centre............................................................................. 02 6285 2946
Website......................................................................................www.capitalpath.com.au
Email ..........................[email protected]capitalpath.com.au
xix
Doctors Service Centre (DSC) 02 6285 9803
Results Enquiries
If you wish to phone us for results or any general enquiry:
Doctors Service Centre (DSC).................................................................. 02 6285 9803
Toll free–outside ACT................................................................................ 1800 807 556
Please supply surname, first name and date of birth of the patient, and the name of the
referring doctor when requesting results.
Alternatively you may wish to speak with a pathologist for interpretation of a result. If so,
please refer to the contact phone numbers on the ‘Contacts Page’.
Confidentiality
To maintain patient confidentiality, the staff in the Doctors Service Centre and other
laboratory departments make every effort to ensure that only authorised recipients are
given results.
Capital Pathology apologises for any inconvenience this may cause.
xx
Collection Centres
Appointments are not required for most tests, however for your patients’ convenience
appointments are available at most of our Collection Centres.
Please phone specific Collection Centre for further details.
Tests that do require an appointment include:
•
•
•
•
Glucose Tolerance Test (GTT)
Helicobacter Breath Test
Holter Monitors
24 Hour Blood Pressure.
ACT
AINSLIE.................................................................................................... 02 6249 1940
100 Wakefield Gardens, Ainslie
BARTON................................................................................................... 02 6273 4505
5/3 Sydney Avenue, Barton
BELCONNEN ........................................................................................... 02 6253 0146
Unit 12 North Point Plaza
8 Chandler Street, Belconnen
CALVARY ................................................................................................. 02 6251 5121
Suite 4, Calvary Clinic,
Haydon Drive, Bruce (rear of Calvary Hospital)
CHARNWOOD ......................................................................................... 02 6258 9197
123 Tillyard Drive, Charnwood
CIVIC ........................................................................................................ 02 6248 0899
34 Marcus Clarke Street, Civic
DEAKIN .................................................................................................... 02 6285 3765
Peter Yorke Building Plaza, Calvary John James Hospital
173 Strickland Crescent, Deakin
DICKSON ................................................................................................. 02 6248 7804
Dickson Professional Centre,
Cnr, Antill and Cowper Streets, Dickson
GARRAN .................................................................................................. 02 6281 7277
Brindabella Specialist Centre,
Dann Close, Garran
GUNGAHLIN ............................................................................................ 02 6242 8328
Unit 132, Hinder Street,Gungahlin
HAWKER .................................................................................................. 02 6255 2241
Unit 7, Birubi Chambers,
Cnr, Beetaloo Street and Hawker Place, Hawker
LANYON................................................................................................... 02 6294 7409
Unit 5/3 Sidney Nolan Street, Condor
xxi
ACT (continued)
MANUKA .................................................................................................. 02 6239 5074
Unit 1/21 Murray Crescent, Manuka
PHILLIP/WODEN ..................................................................................... 02 6293 3860
Suite 11 Corinna Chambers
36–38 Corinna Street
TUGGERANONG ..................................................................................... 02 6293 3860
Unit 5, Cnr, Anketell and Reed Streets, Tuggeranong
WANNIASSA ............................................................................................ 02 6296 1644
Unit 2, Erindale Chambers,
Gratton Court, Erindale Shopping Centre, Wanniassa
NSW
BEGA ....................................................................................................... 02 6492 2049
101 Carp Street, Bega
BERRIDALE ............................................................................................. 02 6452 3150
6 Myack Street, Berridale
COOMA .................................................................................................... 02 6452 3150
190 Sharp Street, Cooma
CROOKWELL .......................................................................................... 02 4843 2555
17 Kialla Road, Crookwell
EDEN........................................................................................................ 02 6496 3508
Curalo Medical Centre
60 Princes Highway, Eden
GOULBURN ..............................................................................................02 4821 4011
127 Bourke Street, Goulburn
JERRABOMBERRA ................................................................................ 02 6299 8642
Unit 12, Jerrabomberra Shopping Village Centre
2 Limestone Drive, Jerrabomberra
JINDABYNE ............................................................................................. 02 6457 2209
Nugget’s Crossing Shopping Centre, Jindabyne
MERIMBULA ............................................................................................ 02 6495 3045
Unit 2/93 Main Street, Merimbula
MERIMBULA (Sapphire Clinic)............................................................... 02 6495 4879
44 Merimbula Drive, Merimbula
PAMBULA ................................................................................................ 02 6494 3024
17 Quondola Street, Pambula
QUEANBEYAN ........................................................................................ 02 6297 3600
1st Floor
23 Antill Street, Queanbeyan
xxii
Specimen Collection
Specimen labelling
A correctly labelled specimen is essential. Persons collecting specimens must positively
identify the patient, and ensure that there are at least two unique identifiers recorded on
the specimen.
1. Patient first and surname.
2. Date of birth ( and/ or medical record number for hospital patients).
A correctly labelled specimen should arrive in the laboratory with a completed pathology
request form that includes the date and time of collection.
There are exacting requirements for blood bank specimens and paperwork.
See Blood Banking.
There are also specific requirements for all antenatal and prenatal specimens.
1.All pre- transfusion and antenatal and perinatal specimens must be labelled with
surname, given name(s), date of birth, date and time of collection
2.The person collecting the blood is to sign or initial the tubes
3.The person collecting the blood is to sign a declaration on the request form
similar to the following:
“I certify that I collected the accompanying sample from the above patient whose
identity was confirmed by enquiry and/or examination of their name band and that
I labelled the sample immediately following collection.”
Home Visits / Hospital Visits
Capital Pathology provides extensive collection services to hospitals and nursing homes.
Staff are also available to collect specimens and perform ECG tracings or Holter Monitors
from patients at home who are unable to visit a Collection Centre. Please phone the
appropriate Collection Centre to arrange a house call for Northside, Southside or Central
locations.
Materials for Specimen Collection
Materials necessary for the collection of pathology specimens can be supplied at no
charge, subject to Medicare guidelines. A form labelled “Request for Pathology Supplies”,
which lists the supplies available, can be obtained from your Capital Pathology courier.
To order supplies, please fill in this form and send in to the laboratory with your courier
or via fax.
xxiii
Courier Services
Routine Courier Services
An extensive courier service is provided throughout our practice areas in ACT and NSW,
seven days a week. Each courier delivers reports and pathology supplies and picks
up specimens. A courier will call upon surgeries regularly, or if preferred, only when
requested. Please contact Client Services Department on 02 6285 9802 to arrange
timings for regular courier pick ups.
Urgent Courier Services
Urgent specimen pick–up is available in emergencies by phoning the Courier Department
on 02 6285 9877.
Specimen Boxes
For convenience, a secure specimen box can be installed at surgeries for specimens
awaiting out of hours pick–up.
Please phone the Client Services Department on 02 6285 9802 to arrange this service.
xxiv
Requesting Pathology Tests
Request Forms
Requests for pathology tests may be written on the surgeries’ own stationery or on Capital
Pathology request forms. The reverse page of pre printed Capital Pathology request forms
details information on our current Collection Centres with opening times and services
offered at each centre. Information on the reverse of form is correct at time of printing.
For request form replacements, please fill in the re–order form included in the request
pad or phone Front Reception on 02 6285 9800.
The following are available:
1.Request forms personalised with the doctor’s name and address.
2. Laser request forms for use in printers.
When completing request forms, please supply the patient’s full name, sex, date of birth,
address and phone number. Relevant clinical history is very helpful to the laboratory,
and this information will be included in the completed report.
Requesting Tests
When ordering tests, please make requests as specific as possible. Certain approved
abbreviations may be used to order tests or groups of tests. Acceptable abbreviations
are published in the Medicare Benefits Schedule book.
Tests do not need to be handwritten, however all requests should be signed by the
requesting practitioner.
Verbal Requests
Requests for pathology tests may be phoned to either the Doctors Service Centre on
02 6285 9803 or your nearest Laboratory. However, to ensure that your patients receive
Medicare Benefits, the laboratory is required to receive signed confirmation of these
tests within 14 days of the verbal request.
The laboratory will send a printed request form, which includes the patient details,
for signing and confirmation of the tests requested to the requesting doctor.
xxv
Requests for a Series / Rule 3 Exemption
Under certain circumstances, Medicare regulations allow a single signed request form
to cover repeat testing over a six-month period. The table below details the maximum
number of repeat tests allowed under these circumstances. Once the maximum number
of tests has been reached, or the six-month period has expired, a new request form for
repeat testing will be required. Please state on the request form “Rule 3 exemption” with
the relevant therapy/condition and the tests requested.
Condition/Treatment
Tests requested
Maximum tests
per request
Anticoagulant therapy
INR
Unlimited
Methotrexate or Leflunomide
therapy
Full Blood Count (FBC)
Basic chemistry items including;
Liver Function Tests (LFT)
Urea, Electrolytes and Creatinine (U&E)
CRP
6
Chemotherapy or
immunosuppressant therapy
Full Blood Count (FBC)
6
Cis-Platinum or
cyclosporin therapy
Urea, Electrolytes and Creatinine (U&E)
6
Lithium therapy
Lithium
6
Gold, penicillamine , clozaril,
ticlopidine hydrochloride or
sulphasalazine therapy
Full Blood Count (FBC)
6
Patients with chronic
renal failure being treated
in a dialysis programme
conducted by a recognised
hospital
Urea, Electrolytes and Creatinine (U&E)
6
Vitamin D therapy
Calcium, Albumin
6
Patients with cancer
receiving biphosphonate
infusions
Calcium, Phosphate, Magnesium,
Urea, Electrolytes and Creatinine
6
(6 month period)
xxvi
Laboratories
Our laboratories have been accredited to ISO 15189 Standard under the accreditation
scheme of the National Association of Testing Authorities and the Royal College of
Pathologists of Australasia (NATA/RCPA). To maintain this recognition, our laboratories are
re-evaluated periodically by NATA to ensure their continued compliance with requirements,
and to check that their standard of operation is being maintained.
ACT
2 Makin Place
Deakin 2600
t: 02 6285 9800
f: 02 6281 1941
South Coast
Unit 7/89 Auckland Plaza
Auckland Street
Bega 2550
t: 02 6492 0013
f: 02 6494 7130
Goulburn
127 Bourke Street
Goulburn 2580
t: 02 4821 4011
f: 02 4821 4224
xxvii
Results 02 6285 9803
Doctors Service Centre.....................................................02 6285 9803
Routine Results
Results are both printed and downloaded continually throughout the day and delivered
routinely to practices throughout ACT and NSW. Grossly abnormal results are phoned
or faxed to the requesting doctor or surgery.
Urgent Results
Requests marked “Urgent” will receive priority processing and results will be phoned
or faxed as soon as available. Please assist us by providing a date and time result
is required.
Results by Fax
As a supplement to hard copies, results can routinely be sent directly to a fax machine.
To arrange for this service, please contact the Doctors Service Centre on 02 6285 9803.
Results by Electronic Download
Results may be downloaded and imported into all major practice management software
systems.
Results may also be viewed via the internet using Webster, our secure result
viewing application.
For further information on results transfer systems and Webster, please phone
IT Services on 02 6285 9860.
xxviii
Reports: paper and ‘paperless’
Patients Details
Your reference number
(if required).
Patients telephone number
(if supplied).
DOB, sex and age.
Episode Information
Lab number in bold.
Referral date included.
Collection date included.
Service date in bold
(date received in laboratory).
Doctor’s Details
Referring doctor or copy
to doctor information
Laboratory/Pathologist
contact information
Patient Details
Clinical history (if supplied).
Result
Standard format for cumulative
results.
Reference ranges given.
Abnormal results bolded
and flagged.
Appropriate results
information
Laboratory Information
Test code for result shown on
this page.
Date of service
Plus recipient initial.
Pathologist Initials
Indicates number of pages
and page sequence in report
Preferences of report style are very much an individual choice, and will vary from surgery
to surgery. Capital Pathology provides several options for customising reports to suit the
requirements of different doctors and other clients.
These report options are available using either A4 or A5 paper size. Please note that a
combination of A4 and A5 is not available.
For customising of your reports, please contact the Client Services Department on
02 6285 9802.
“Interims Only”
This allows for reports to be issued as results are available.
xxix
“Finals Only”
“Finals Only” reports are produced when all results on a single patient episode number
have been completed (excluding Histology, Cytology and tests sent to other laboratories).
Only one report is produced when the final test on the patient’s episode is completed.
The advantage of this system is the reduction in the number of reports produced requiring
filing. However, the report is not generated until the “longest to complete test” is finalised.
This type of report is suitable for surgeries that have an electronic download of their
pathology reports, so the possibly increased turnaround time of the final printed report will
not be significant.
Accumulated Interims
Reports are produced as each test is completed, and include all other reported tests from
that particular episode on the report. This means that when the final test is completed, all
tests will be included on the report (excluding Histology, Cytology and tests sent to other
laboratories). As progressive reports are available as tests are completed, turnaround time
of reports is not compromised. However, additional reports and therefore extra paper is
produced.
Interims / Finals
Interim reports are produced as each test is completed. A “Final” report is printed once
all tests have been completed and includes all tests on the episode. This will result in
individual tests being available with no delay in turnaround time and a consolidated report,
with all patient results (excluding Histology, Cytology and tests sent to other laboratories),
also being produced.
Cumulatives
Cumulative reports are available for certain tests. These list past and current results on the
one report and are designed to facilitate monitoring by the practitioner. The most recent
results appear on the right hand side of the report.
Duplex (Double–Sided)
Single–sided reports are the standard report format produced. As surgeries use a wide
range of record filing systems, a choice of either single or double–sided reports (duplex)
is available.
Paperless
Individual doctors, or complete surgeries may decide they no longer wish to receive paper
reports and only use electronic copies. If you wish to arrange this, please contact our
Client Services Department on 02 6285 9802.
Copies of Reports
If extra copies of reports are required, please indicate the name and address of the
recipient in the “copy to” space on the request form. This space may also be used to
indicate if the surgery wishes to receive additional copies. If multiple copies of reports are
always required, this can be arranged by phoning the Client Services Department
on 02 6285 9802.
xxx
Billing Policy
Capital Pathology is a private pathology practice without direct government subsidy.
Medicare cutbacks over the past decades have meant that average rebates for testing are
now 10% less in actual terms and 60% less in real terms than they were in 1985. Over the
past decade there have been no Medicare rebate increases, but two sizable cuts occurred
in 2008 and 2009. We are, however, aware of the ever increasing fee burden for our
patients, so have put in place mechanisms to minimise patient costs.
For out-patient testing, our practice fees are generally those recommended by the AMA,
which we currently cap, so that for each episode of testing ‘gap’ out of pocket costs are
limited to $50. This means your patients usually pay no more than $50 for an episode of
Medicare rebateable testing after claiming their rebate from Medicare.
Patient Accounts
We have a range of brochures outlining our billing policy for both inpatients and
outpatients that you may find of benefit. Please contact us if you wish to obtain copies
for your practice.
Payment for accounts may be made by either paying the account directly to Capital
Pathology first and then claiming the rebate from Medicare, or by claiming from Medicare
first and the patient then forwarding the rebate cheque to us together with any additional
payment. Details of payment options are included with the account.
For any further enquiries please contact our Accounts Department on 02 6285 9888
or Client Services Department on 02 6285 9802. Out-patient Services
We routinely bulk bill Medicare eligible patients who hold the following cards:
•
•
•
Pensioner cards
Healthcare cards
Veteran Affairs.
We also bulk bill:
•
•
Children under 16
Nursing home patients.
And as directed by the referring doctor.
xxxi
In-patient Services
In-patient pathology tests are performed for patients who are:
•
•
A private patient in a private hospital or approved day hospital facility
A private patient in a recognised (public) hospital.
Fees for patients who have eligible cover with a health fund that has a no-gap agreement
with Capital Pathology.
Patients who have eligible private health insurance with a Health Fund that has a no-gap
agreement with Capital Pathology may have their account billed directly to Medicare
Australia and the private health fund. They will not incur any additional out of pocket
expenses for tests that are eligible for Medicare rebates. For information on whether
specific health funds are covered by such an agreement, please contact our Accounts
Department.
Fees for uninsured patients and those covered by a health fund that does NOT have an
agreement with Capital Pathology.
Patients who are uninsured or who do not have eligible cover with a health fund will
receive an account for their pathology tests. Our fees for in-patient testing are generally
at, or below, those recommended by the AMA. When the account is paid, the patient may
present the receipt to Medicare Australia and their private health fund to claim their rebate.
Medicare Rebate Eligibility
Please note that some pathology testing is not eligible for a Medicare rebate. Patients who
present to our Collection Centres are informed of the approximate cost involved prior to
testing taking place.
Pathology tests for the following circumstances are also not eligible for a Medicare rebate:
•
•
•
•
•
•
employment immigration insurance superannuation Work Cover
visa applications.
In some cases, tests may be sent to a reference laboratory who may send out an
individual account. Details are available from individual Collection Centres or the
Specimen Reception Department on 02 6285 9873.
Corporate accounts may be initiated where testing is independent of Health Insurance
Commission guidelines. Please contact the Collection Manager on 02 6285 9881 or
access Corporate Services at our website on www.capitalpath.com.au
xxxii
The Coning System
In 1995 Federal Government budget changes saw the introduction of a scheme in
which Medicare payments cut out after three pathology items on any single episode
ordered by General Practitioners. This change should not have restricted the ordering of
pathology tests in any way (i.e. there is no need for doctors to change their usual pattern
or diagnostic approach). With the greater complexity of modern medicine and the ageing
population, the number of patient episodes with more than three requested items has
been steadily increasing. However, we absorb the cost of tests beyond the three items so
the patient suffers no extra out-of-pocket expenses.
Pap smears are not included in the coning scheme.
Patient Episode Initiation Fees (P.E.I.)
This fee was introduced by the Health Insurance Commission in 1992 in conjunction with
reduced Medicare benefits for certain pathology tests.
An initiation fee is designed to:
1.Represent costs other than those directly involved in the actual test procedure and
includes transporting specimens to the laboratory, collecting samples, forwarding the
completed report to the referring doctor and administration costs.
2. Describe to the Health Insurance Commission how the pathology service originated.
e.g. Collection Centre, doctor collection or hospital collection.
A pathology provider can charge only one initiation fee per referral unless a Rule 3
Exemption applies. Please see section entitled ‘Requests for a Series / Rule 3 Exemption’
under “Requesting Pathology Services”.
The fee is claimable from Medicare along with the fee/s for the actual pathology test/s.
xxxiii
Education
Capital Pathology has a commitment to provide ongoing education to medical
practitioners, practice and nursing staff.
Specialists
Our specialist Pathologists run regular educational sessions throughout the year with
various groups of specialists to review specific cases of interest.
General Practitioners
We offer regular educational workshops throughout the year on various interesting and
topical subjects.
Practice and Nursing Staff
Workshops are held regularly by Capital Pathology Senior Scientists throughout the year.
For more information on any of these educational sessions please contact the Client
Services Department on 02 6285 9802 or access the website on www.capitalpath.com.au
xxxiv
Publications and website www.capitalpath.com.au
Capital Pathology aims to provide up to date information on the range of services provided
by our practice, plus publish articles relating to new developments and changes in
pathology and health in general.
Patient Information Sheets
Patient Information Sheets are available regarding a number of common test procedures
and ‘What is pathology?’ These are available on our website, from individual Collection
Centres or by contacting our Stores Department on 02 6285 9813.
Billing brochures
Information regarding billing for both inpatients and outpatients are available on request.
Please contact Client Services Department on 02 6285 9805 if you would like copies.
Doctor’s Newsletters and Capital Letter
These publications are printed regularly throughout the year and are available on our
website, or by contacting the Client Services Department on 02 6285 9802.
Doctor’s Handbook
Copies are available in hard copy, CD or on the website. Please contact the Client
Services Department on 02 6285 9802 if you would like copies in either format.
Guides
Laminated guides are available on a range of subjects including:
Vacutainer Guide
Swab Guide
Paediatric Tube Guide
Calenders
Collection Centre Listings
Reference Ranges
Test Abbreviations
Please contact the Client Services Department on 02 6285 9802 if you would like copies
of any of these.
Kid’s Courage Pack’s
We understand that having a specimen taken can be a stressful procedure, particularly for
our younger patients. As such we have developed a ‘Kid’s Courage Pack’ which contains
a number of activities to help children during this time. These are available at all our
Collection Centres.
Website
Our website has information updated regularly and is accessible on www.capitalpath.com.au
xxxv
Other Services
Other services provided by Capital Pathology include:
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Biological Indicator monitoring
Blood Bank and transfusion service
Bone Marrow collections
Drug Testing
Corporate Medical Screening and Vaccinations
ECG Recording and Interpretation by specialist Cardiologists
Fine Needle Aspiration
Frozen Section diagnosis
Genetic Testing
Helicobacter Pylori breath testing
Holter Monitor service and reporting by specialist Cardiologists
Infection Control Advice for hospitals, nursing homes and surgeries
Mantoux Tests (Tuberculin Sensitivity Test)
Pap Smear Reminder service, including follow-up letters for abnormal pap
smears and statistical evaluation of pap smears
Spirometry
Water testing
24 Hour Blood Pressure Monitors.
For further information on any of these services, please contact the Client Services
Department on 02 6285 9802.
xxxvi
Corporate Services
Capital Pathology is available to assist businesses assess known or suspected workplace
risk factors that may contribute to ill-health and thus potential loss of productivity.
Our dedicated Corporate Services Team are able to perform a variety of tests and
procedures both on and off site as part of the assessment of workers and workplaces.
The following tests are available:
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Corporate health testing
Insurance pathology
Insurance paramedicals
HIV antibody testing
Hepatitis screening
Vaccination programmes including hepatitis A & B, tetanus and influenza
Audiometry- hearing tests
Spirometry- lung function
Cardiologist reported ECG services including 24hr ambulatory
Occupational biological monitoring
Occupational allergy testing
Heavy metals testing
Trace metals toxicology
Drug and alcohol testing (AS4308)
Chemical exposure monitoring
Pesticide exposure monitoring
Water testing
Therapuetic & drug trial monitoring
Genetic studies
Zoonoses – Q fever.
Please contact Collection Manager on 02 6285 9881
or email [email protected] for further information
xxxvii
Capital Pathology Handbook – Interpretation of Laboratory Tests
A
A
Acetylcholine Receptor Antibody
Specimen:
Serum – Gel
Reference Range: Not detected
This is a useful indicator in the diagnosis of Myasthenia gravis, with receptor
antibodies detected in more than 90% of patients with generalised myasthenia.
Acid–Base Status
See Bicarbonate
Acid–Fast Bacilli (AFB)
SeeMantoux Test
Tuberculosis
ACTH (Adrenocorticotrophic Hormone)
Specimen:Plasma – EDTA
Spin, separate and freeze plasma immediately.
Reference Range: Supplied with report
Note: special collection requirements apply. It is preferable to send patient to
collection centre.
Actinomyces
An anaerobic gram–positive filamentous bacillus found as a normal commensal in the
mouth and gut. The most commonly encountered species is A.israelii. Actinomyces species
are susceptible to penicillin and doxycycline. Most isolates are resistant to metronidazole.
The most common form of infection is cervicofacial infection characteristically occurring
in association with poor oral hygiene and tooth abscesses or decay. Thoracic and
gastrointestinal infections also occur. All these deep–seated infections may declare
themselves by developing draining sinuses.
Actinomyces species also grow in association with IUCDs (intrauterine contraceptive
devices). They may be detected in a cervical smear or by culture of a removed IUCD
in an asymptomatic woman. Although pelvic actinomycosis has been reported in
association with IUCD use, the individual risk of infection with a colonised IUCD is small.
Activated Protein C Resistance (APCr)
Specimen:Plasma – Sodium Citrate x 2
If being performed as part of a thrombophilia screen
collect sodium citrate x 6
Sodium Citrate tubes must be filled to capacity.
Reference Range: Supplied with report
Activated protein C resistance occurs in patients with the Factor V mutation known
as Factor V Leiden (FVQ506). This mutation is present in 5% of a normal Caucasian
population. It is the commonest of the recognised thrombophilia markers. It is
detected in 16–20% of consecutive patients presenting with thromboembolism but the
frequency is higher in selected patient groups. Although it increases the thrombotic
risk 5–10 fold, it is generally regarded as a relatively weak risk factor.
APCr is measured using an APTT ratio with and without activated protein C. The
normal ratio is 2.4–4.0. Heterozygous positive patients with the Factor V mutation
have ratios of 1.6 or less. Homozygosity, which is much less common, is associated
with a higher thrombotic risk and a lower APCr.
See:
Factor V Leiden
Acute Leukaemias
The diagnosis is usually obvious because of blast cells in the blood film, raised total
white count, anaemia, thrombocytopenia and neutropenia, but one or more of these
features can be absent at the time of diagnosis.
Acute Lymphoblastic Leukaemias (ALL)
The more common childhood leukaemia. Prognosis is relatively good in children with
80–90% remission rates. Remission rates are lower in adults.
Acute Myeloid Leukaemias (AML)
The more common leukaemia in adults. AML may arise de novo or through
transformation of chronic myeloid leukaemia or myelodysplastic syndromes.
Transformed AML responds poorly to treatment.
In de novo AML, first remission is achieved in 70–80% of patients with 30–40% long
term survivors when treated with chemotherapy alone. With allogeneic bone marrow
transplantation for patients aged less than 55 years there is a 50–60% five year
survival.
The M3 subtype, acute promyelocytic leukaemia (APML), is treated with ATRA
(a retinoic acid derivative) initially, to promote cell differentiation, followed by
chemotherapy. This gives 85–90% remission rate with 80% 12–month disease free
survival.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Acute Phase Proteins (reactants)
Specimen:
A
Serum – Gel and EDTA
Reference Range: Supplied with report
Following inflammation or tissue destruction there is an increase in the so–called
acute phase proteins with the development of a typical pattern on serum protein
electrophoresis.
Analyte
Rise
Response times
CRP
Up to 1000x
6–10h
Alpha 1 antitrypsin
2–4x
24–48h
Haptoglobin
2–4x
24–48h
Fibrinogen
2–4x
24–48h
Ceruloplasmin
1.5–2x
48–72h
C3
1.5–2x
48–72h
C4
1.5–2x
48–72h
Adenovirus Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Adrenal Antibodies
Specimen:
Serum – Gel
Reference Range: Not detected
Adrenal antibodies are present in 80% of autoimmune Addison’s disease.
Adrenaline
SeeCatecholamines
Adrenal Function
Suspect?
Test
Addison’s disease
Cortisol, ACTH, Adrenal cortex Abs, Synacthen stimulation
test – due to risk of anaphylaxis, the Synacthen test is
only to be performed under medical supervision with
access to full resuscitation facilities
Congenital adrenal hyperplasia
17–OH Progesterone
Cushing’s Disease
AM/PM cortisol, DHEAS, Prolonged dexamethasone
suppression test, Short dexamethasone suppression test,
Testosterone, U–Free cortisol
Phaechromocytoma
24-hour U–Catecholamines
Aeromonas
Aeromonads are widely distributed in stagnant or flowing fresh water and in soil.
Human infections include:
•
•
•
•
Cellulitis or wounds contaminated with soil or water
Acute diarrhoeal disease–worldwide, affecting any age
Septicaemia
Other infections, including the urinary tract.
Aeromonads are mostly resistant to the penicillins but are often susceptible to
trimethoprim/ sulphamethoxazole and quinolones, e.g. ciprofloxacin. Nalidixic acid
or nitrofurantoin may be used for urinary tract infection.
AFP (Alpha–fetoprotein)
Specimen:
Serum – Gel
Reference Range: Age related reference ranges supplied with report
AFP has two uses:
•
•
Tumour marker–large elevations (> 100) can be found in hepatomas and testicular
tumours and are used for monitoring treatment and in diagnosis.
Smaller non–diagnostic elevations are found in many forms of liver disease and
malignancy.
Screening for neural tube defects and Down’s syndrome
See
Pre-Natal Testing
Alaninine Transaminase or Amino Transferase (ALT)
Specimen:
Serum – Gel
Reference Range: Adult 5–40 U/L
Age related reference ranges supplied with report.
See
Liver Function Test / Interpretation
Capital Pathology Handbook – Interpretation of Laboratory Tests
Albumin
Specimen:
Serum – Gel
Reference Range: Adult 35–50 g/L
Decreases:
•
•
•
•
•
Acute or chronic inflammatory illness
Urinary protein loss
Advanced liver disease
Gastrointestinal loss
Severe malnutrition.
Increases are usually due to dehydration.
Albumin Excretion Rate (AER)
Specimen:Urine – Timed overnight collection
24 hour collection (nil preservative).
Urine results expressed as Albumin Excretion Rate (AER).
Reference Range: < 20 ug/min normal
20–200 ug/min indicates microalbuminuria
> 200 ug/min indicates clinical proteinuria
Alcohol
Specimen:
egal Whole blood – EDTA or Flouride Oxalate or Heparinised
L
blood with tamper proof seal.
Special collection requirements apply, please contact Doctors
Service Centre for further information.
Non-Legal Whole blood – Flouride Oxalate
Reference Range: Supplied with report
Aldosterone
Specimen:Plasma – EDTA
Urine – 24 hour (nil preservative).
Reference Range: Supplied with report
A
Alkaline Phosphatase (ALP)
Specimen:
Serum – Gel
Reference ranges vary with age and sex, with an increase during the teenage growing
years and also with a gradual increase with advancing age. See individual test reports
for specific reference ranges.
Reference range in adults 25 to 45 years
Males
35–110 U/L
Females
20–105 U/L
The bone isoenzyme comes from active osteoblasts, hence high levels during
childhood, pubertal growth spurt, healing fractures.
In the liver, ALP is found in biliary canaliculi, increased production being stimulated by
biliary obstruction and to a lesser extent other liver pathology.
Causes of hyperphosphatasaemia can be group under three main headings:
Liver disease
ALP of liver origin is usually identified by an accompanying rise in GGT which is also
an “obstructive” enzyme. AST and ALT are usually also elevated to varying extents in
liver disease.
For further discussion see Liver Function Test / Interpretation
Bone and other non–liver disease
• Paget’s diseases of bone
• Malignant disease, secondary or primary
• Hyperparathyroidism
• Rickets/osteomalacia
• Chronic renal failure
• Healing fractures
• Thyrotoxicosis
• Rheumatoid arthritis
• Intestinal disease, e.g. Crohn’s, ulcerative colitis.
Benign isolated elevations of ALP
• Growth in childhood, particularly the pubertal growth spurt when levels can be up
to 650 U/L.
• Third trimester of pregnancy, up to 400 U/L.
• Isolated elevations of ALP are common in the elderly and usually attributed to
foci of subclinical Paget’s disease provided there is no evidence of malignancy.
Another possibility is osteomalacia due to vitamin D deficiency.
• In transient hyperphosphatasaemia of infancy and early childhood, ALP can go up
to 3000 U/L for up to 3 months. Other liver enzymes remain unaffected. Aetiology
remains obscure but viral infection is a possibility.
• Familial benign hyperphosphatasaemia shows as a raised ALP throughout life.
Capital Pathology Handbook – Interpretation of Laboratory Tests
EVALUATION OF AN ISOLATED SERUM ALKALINE PHOSPHATASE (ALP) ELEVATION
Possible causes
Further investigations
Consider
Pregnancy (increased placental isoenzyme)
Age < 20 years (may be physiological)
Due to the skewed distribution of ALP, levels of up to 400 IU/L are not uncommon in
growth periods. Levels higher than this have also been reported.
ALP Isoenzymes
Predominant bone Bone scan (if indicated)
Paget’s disease
Healing fractures
Renal osteodystrophy
Hyperparathyroidism
Hyperthyroidism
Malignancy
(prostate, breast, kidney, myeloma, lymphoma, etc.)
Osteomalacia/Rickets
(due to Vitamin D deficiency or resistance)
Placental (Regan isoenzyme)
Malignancy (bronchus, ovary, pancreas)
Predominant liver
Cholestatic liver disease
THI pattern (child)
Transient Hyperphosphatasaemia of Infancy.
Typical age < 5 years; very high ALP (> 700).
May follow a viral illness.
Benign and asymptomatic.
High ALP persists for 8–12 weeks.
A
SERUM ALKALINE PHOSPHATASE (ALP) ELEVATION
SuggestedSerum
scheme
for evaluation
of a high
serum
ALP
Alkaline
Phosphatase
(ALP)
elevation
ALP High 2
Suggested scheme for evaluation of a high serum ALP
Consider:
• Pregnancy
• Age < 20 years1
(may be physiological)
Estimate Bilirubin,
ALT, GGT
Bilirubin
High
Normal
ALT
High
Cholestatic liver disease:
Extrahepatic obstruction
Intrahepatic obstruction
Hepatitis
Alcoholic liver disease
Primary biliary cirrhosis
Sclerosing cholangitis
Ascending cholangitis
Post-operative cholestasis
Gram negative bacteraemia
Drug toxicity
Oestrogens
Anabolic steroids
Antipsychotic drugs
Synthetic penicillins
Erythromycin
Gold
Captopril
Normal
GGT
Normal
High
Investigative protocols Elevated ALP Ix
Isolated elevated ALP
Mixed hepatocellular
and cholestatic disease
Chronic active hepatitis
Space-occupying lesion 2
Cirrhosis
Drugs (see cholestatic list)
ALP Isoenzymes
? Cholestatic liver disease
? Two processes i.e.
1. Liver enzyme induction 3
2. Bone disease
‘Placental’
(Regan isoenzyme)
Malignancy
Bronchus, Ovary, Pancreas
Predominant liver
Cholestatic liver disease
(see above)
‘THI’ Pattern (child)
Predominant bone
Bone scan
(if indicated)
Transient Hyperphosphatasaemia of Infancy 4
Paget’s disease
Malignancy 5
Healing fractures
Renal osteodystrophy
Osteomalacia/Rickets 6
Hyperparathyroidism
Hyperthyroidism
1. Due to the skewed distribution of ALP, levels up to 400 IU/L
are not uncommon during growth periods—higher levels
have also been reported.
2. Malignancy (primary, secondary), abcess, cyst.
3. Alcohol, drugs (phenytoin, warfarin, benzodiazepines,
tricyclics), obesity, diabetes mellitus, hypertriglyceridaemia.
4. Typically age < 5 years; very high ALP (> 700); may follow
a viral illness. Benign and asymptomatic; high ALP persists
for 8–12 weeks.
5. Prostate, breast, kidney, myeloma, lymphoma, etc.
6. Due to Vitamin D deficiency or resistance.
90 Sullivan Nicolaides Pathology
Capital Pathology Handbook – Interpretation of Laboratory Tests
Alkaline Phosphatase Isoenzymes
Specimen:
Serum – Gel
Assessment of other laboratory tests and the clinical picture will usually explain
elevations of ALP without resorting to isoenzyme fractionation–which usually provides
a clear–cut answer only when the answer is already obvious.
A useful generalisation is that, where both ALP and GGT are raised, liver is the
source. When GGT is normal, the raised ALP is probably from bone.
Occasionally, however, the answer is not obvious. Heat stability is a method of
isoenzyme analysis, the ALP being measured before and after 10 mins incubation
at 56˚C.
%ALP remaining
after incubation
Likely origin of raised ALP
< 20%
mainly bone
25–55%
mainly liver and/or intestine
> 90%
placenta
Allergy
Tests used in the diagnosis of IgE-mediated allergy
Once evidence of hyperreactivity is established, allergy testing plays a significant
role in the diagnosis of allergy. It is important to note that an allergy test cannot have
100% specificity and sensitivity. Changing the cut-off level to improve one parameter
will often worsen the other parameter. Hence cut-off levels are set to maximise both
sensitivity and specificity.
Total IgE
The majority of patients with allergies have elevated total IgE levels. Higher serum
IgE levels are seen in hyperreactivity diseases in which larger parts of skin and/or
mucosa are involved. Patients with atopic dermatitis tend to have higher IgE levels
than asthmatics who, in turn, have higher levels than patients with allergic rhinitis. In
the absence of an increased IgE, further investigations for IgE allergy are likely to be
unproductive. Therefore, serum IgE level can be used as a simple assessment of a
patient’s allergic profile.
Allergen-specific IgE
Specific IgE may be determined from a range of allergens using a non-isotopic
variation of the radioallergosorbent test (RAST). RAST results are not affected by
antihistamine or corticosteroid intake. The sensitivity and specificity of the RAST
varies with the allergen and the allergy site. For inhalant allergies, the sensitivity of the
RAST is 60–80% and the specificity is higher than that of skin tests, often as high
as 90%. Please ring our Doctor’s Service Centre (DSC) with any further enquiries.
Skin tests
Skin tests for a range of allergens are usually performed on the volar aspect of the
forearm. Antihistamine intake may cause false negative results.
As with the RAST, skin test sensitivity and specificity varies with the allergen and
allergy site. Generally, the sensitivity of skin tests is higher than that of the RAST.
However, the specificity of the test is often lower (70–80%).
A
Alopecia
The common age and gender related variant does not warrant laboratory investigation
but specific aetiologies to be considered include:
•
•
•
•
•
•
•
•
Hypothyroidism
Hyperthyroidism
Drugs: cytotoxics, lithium
Dermatological pathology (see Skin Biopsy)
Any severe illness
Zinc deficiency
Warfarin hypersensitivity (rare)
Polycystic ovary syndrome.
When associated with a severe but correctable illness, the alopecia is likely to be
transient.
Alpha–1–antitrypsin
Specimen:Serum – Gel
Genotype Whole blood – EDTA.
Reference ranges: Supplied with report
AAT deficiency in its severe forms predisposes to early onset emphysema and juvenile
cirrhosis and should be sought in these situations.
Sometimes the deficiency is detected as an absent or markedly reduced alpha–1 band
on routine serum protein electrophoresis.
Genetically there is one normal gene, M, and two abnormals, S and Z
Genotype
Prevalence (%)
Reduction
AAT level (%)
MM
88
0
MS
7
20
MZ
4
40
SS
1
40
SZ
0.1
70
ZZ
0.03
90
ZZ and, to a lesser extent, SZ are associated with emphysema and cirrhosis, and both
smoking and alcohol should be strongly discouraged. The importance of MS, MZ and
SS is uncertain but avoiding smoking is a wise precaution.
Tests for AAT deficiency:
Protein electrophoresis — a crude screen test but an absent or reduced alpha–1 band,
or a band of slow mobility, is often the first clue to AAT deficiency.
AAT quantitation–a result of < 1.0 g/L is suspicious and requires phenotyping which
gives the best measures of risk.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Aluminium
Specimen:Whole blood – Trace element tube
Urine – 24-hour urine (nil preservative) or 50 mL random urine.
Reference Range: Supplied with report
A
Amenorrhoea
EVALUATION ALGORITHM
Amenorrhea/Irregular
periods
Pregnancy
Thyroid
dysfunction
Exclude
Excercise
induced
Primary
amenorrhea*
Chronic
illness
Anorexia
Nervosa
OCP
Serum: FSH, LH
Oestradiol (E2)
Prolactin (PRL)
FSH:N
LH:↑
E2:N
? Polycystic
ovaries
FSH:↑
LH:↑
E2:↓
Ovarian Scan
Ovarian failure
? menopause
Hypothalamic/pituitary
dysfunction
Consider:Stress-related
anovulation †
Anorexia/dieting
Severe excercise
Psychosocial
stress
FSH:↓
LH:↓
E2:↓
PRL:↑
Hyperprolactinanaemia
* Chromosome abnormalities/ovarian failure more common in women who have never had a spontaneous period.
† FSH:LH ratio often > 1.0
Capital Pathology Handbook – Interpretation of Laboratory Tests
Amiodarone
Specimen:Whole blood – Lithium heparin tube
Wait at least 3 months after commencing therapy before sample
collection, unless toxicity is suspected.
Please provide the date that therapy started.
Collection just before next dose (at least 8 hours post dose).
Reference Range: Supplied with report
Amitriptyline
Specimen:Plasma – Lithium heparin
Trough level is taken before next dose (within one hour).
Reference Range: Supplied with report
Ammonia
Specimen:Plasma – EDTA
Spin and separate plasma within 20 minutes of collection and test
to be performed within 2 hours of collection.
Reference Range: Supplied with report
Amphetamines (Drug Screen)
Specimen:
Random urine
Reference Range: Not detected
See
Drug Screen
A
Amylase
Serum
Specimen:
Serum – Gel
Reference Range: 20–100 U/L
Amylase, found mainly in pancreas and salivary glands, is used in the differentiation
of acute pancreatitis from other causes of the acute abdomen.
In acute pancreatitis, amylase typically rises above 400 U/L, returning to normal in
about 4 days.
The enzyme pattern is inconsistent, however, and failure to detect an elevation does
not preclude the diagnosis even when there is severe infarction.
Acute peritonitis, causing inflammation of the serosal surfaces of the pancreas and
other organs, can elevate amylase but usually not above 400 U/L.
Other causes of hyperamylasaemia include mumps, diabetic ketoacidosis, biliary tract
disease, renal insufficiency, shock, some drugs (particularly narcotics), hypoxia, pelvic
infection, macroamylasaemia.
Chronic pancreatitis does not raise amylase except sometimes during acute
exacerbations.
In macroamylasaemia, as in other macromolecular enzyme variants, a consistently
elevated enzyme level is found in a well person. A definitive diagnosis can be made
using the amylase/creatinine clearance ratio (ACCR).
ACCR(%)=
urine amylase
serum amylase
x
serum creatinine
urine creatinine
x 100
In macroamylasaemia, the ACCR is < 2%. The usual reference range is 2–5%.
Urine
Specimen:
Random or 24–hour urine (nil preservative)
Reference Range: Supplied with report
Urine amylase is elevated in all conditions where serum amylase is raised except
renal failure and macroamylasaemia. It is not often used except perhaps in the
diagnosis of macroamylasaemia where the ACCR is calculated.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Anaemia
A
Anaemia Investigation
An Approach to the Investigation of a Decreased Haemoglobin
Hb/Hct
Exclude blood loss
?stool, urine, sputum
MCV/MCH/MCHC/RDW
film, reticulocyte count
↓ MCV ↓ MCHC
Microcytic hypochromic
Iron deficiency
Thalassaemia trait
Anaemia of chronic
disease
Congenital sideroblastic
anaemia
↑ MCV
Macrocytic
B12/folate deficiency
Myelodysplastic syndrome
(including acquired
sideroblastic anaemia)
Drug induced
Non-megaloblastic
Liver disease
Hypothyroidism
Normal MCV
Normochromic, Normocytic
↓ Reticulocyte count
↓ Bone marrow production
Primary bone marrow failure
e.g. Aplastic anaemia
Acquired red cell aplasia
Secondary bone marrow failure
e.g. Uraemia
Endocrine
Anaemia of chronic disease
HIV
Metastasis
Myelofibrosis
↑ Reticulocyte count
↑ Bone marrow production
Haemolysis
Splenic sequestration
Blood loss
Anaerobic infections
Anaerobic organisms, which are widely distributed in soil and as part of the normal
flora of mouth, gut and vagina, grow readily in deeply–placed abscesses in the
abdomen, wounds, pleural cavities, brain, oropharynx and also in peritonitis or
endometritis. Foul–smelling discharge indicates anaerobic infection.
Routine cultures do not grow anaerobes which require special media and an oxygen–
reduced environment. Anaerobic cultures are set up only on specific request or when
clinical details indicate the need for them. Swabs are not the ideal specimen, tissue
obtained at debridement being the ideal. Aspirated pus or fluid is best collected
and transported in a syringe which should not be refrigerated. IUCDs (intrauterine
contraceptive devices) are routinely cultured for Actinomyces.
Common anaerobic species are Bacteroides, Fusobacterium, Actinomyces and
Clostridium.
Most of the anaerobes we isolate will be susceptible to amoxycillin–clavulanate,
clindamycin and metronidazole.
ANCA (Antineutrophil Cytoplasmic Antibodies)
Specimen:
Serum – Gel
Reference Range: Not detected
These autoantibodies are directed against the cytoplasm of neutrophils.
Androstenedione
Specimen:
Serum – Gel
Reference Range: Supplied with the report
Androstenedione is secreted by the adrenal cortex, testis and ovary and is a precursor
of testosterone and oestrone. In premenopausal women, the adrenal cortex and
ovaries contribute equally but after the menopause androstenedione is almost entirely
of adrenal origin.
In hirsutism and polycystic ovary syndrome it can be elevated but testosterone is the
preferred test.
Androstenedione can be markedly elevated in adrenal hyperplasia and is used in the
investigation of virilism.
Angiotensin Converting Enzyme (ACE)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Although ACE levels can be used in the diagnosis of sarcoidosis (two thirds of patients
with active disease have elevated ACE), elevations are also found in other chronic
inflammatory disorders, giving the test poor specificity.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Anion Gap
Specimen:
Serum – Gel
Reference Range: 10–22 mmol/L
The anion gap is a simple calculation which, if increased above 16 mmol/L, gives
a crude (very crude) indication of metabolic acidosis as in salicylate or methanol
poisoning, ketoacidosis, lactic acidosis, renal acidosis.
The calculation and its derivation are as follows:
In serum, total anions = total cations
C1–+ HCO3 + acid anions (the anion gap) = Na+ (the major cation)
Anion gap = Na+ (C1–+ HCO3)
For example a patient with salicylate poisoning might have
Na+ = 146
C1–= 100
HCO3 = 16
The anion gap, at 30 mmol/L is increased, with acidic salicylate anions being the
cause of the increased gap.
Ankylosing Spondylitis
A seronegative spondyloarthropathy causing low back pain and reduced spinal
mobility due to sacroileitis and spondylitis. Typically it occurs in younger males with
95% positivity for HLA B27.
Anorexia Nervosa and Bulimia
Laboratory abnormalities include:
•
•
•
•
•
•
Hypokalaemia is common in both conditions and can be profound (< 2.0) in
bulimia due to the combination of vomiting and use of laxatives. In this situation
there may be a metabolic alkalosis with raised serum bicarbonate.
Anaemia (normocytic).
Low FSH/LH accompanying the amenorrhoea which is almost invariable in anorexia.
Hypoalbuminaemia, particularly when there is oedema.
Hypocalcaemia is uncommon but is found occasionally.
Cortisol may be elevated.
Other wasting disorders need to be excluded:
•
•
•
glucose to exclude diabetes
TSH/T4 to exclude hyperthyroidism
Addison’s disease causes wasting but is rare in young women under the age of 25
which is the common age group for eating disorders.
A
Antenatal Testing
Routine antenatal care involves looking for several diseases or maternal conditions
that can affect either mother or baby.
The recommended tests can be considered at different stages throughout pregnancy.
Before 20 weeks:
•
•
•
•
•
Full Blood Count (FBC)
Blood group
Red cell antibody screen
Rubella antibodies
Hepatitis B (HbsAg)
•
•
•
•
•
Hepatitis C (HCV) antibody
HIV antibody
Syphilis serology
Urine microscopy and culture
Cervical cytology if indicated.
At 26–28 weeks:
•
•
Red cell antibody screen
Glucose load.
At 30–36 weeks:
•
•
Full Blood Count
Group B streptococcus screening may be indicated.
If high risk for infection:
•
•
Urine or genital swab for Chlamydia trachomatis, gonococcus PCR
Viral swab for herpes simplex PCR.
Additional tests if appropriate:
•
•
1st trimester screening
2nd trimester screening.
If routine antenatal screening returns a positive result, or if the patient is felt to be at
risk for any other reason, further prenatal testing may be required.
It is preferable to specify the individual test required. However, if a request is received
for ‘antenatal serology’ or ‘antenatal screen’ then the following tests will be performed:
Full Blood Count (FBC), Blood group and Red cell antibody screen, Rubella
antibodies, Hepatitis B surface antigen (HbsAg) and syphilis serology. Please note:
Hepatitis C and HIV serology tests require patient counselling and consent, and need
to be individually specified on the request form.
Please note labelling requirements.
1.All pre- transfusion and antenatal and perinatal specimens must be labelled
with surname, given name(s), date of birth, date and time of collection
2.The person collecting the blood is to sign or initial the tubes
3.The person collecting the blood is to sign a declaration on the request form
similar to the following:
“I certify that I collected the accompanying sample from the above patient whose
identity was confirmed by enquiry and/or examination of their name band and that
I labelled the sample immediately following collection.”
See
Pre-Natal Testing
Capital Pathology Handbook – Interpretation of Laboratory Tests
Antibody Screen (Red Cell)
Specimen:
Whole Blood EDTA
See
Coomb’s test
A
Anticonvulsants
Trough levels are usually preferred. Trough levels are taken just before the next dose.
Peak levels may be collected 3–8 hours after the last dose. Timing of peak levels
varies with the individual drug being measured. Carbamazepine peak level 3 hours
post dose, Ethosuxamide peak level 2–4 hours post dose, Phenobarbitone peak level
1–3 hours post dose, Phenytoin peak level 4–7 hours post dose, Primidone peak level
1–3 hours post dose.
Drug
Type of Serum
specimen
Half–life
(hours)
Carbamazepine(Tegretol)
Gel
20
Clobazam(Frisium)
Plasma – Lithium heparin
Clonazepam(Rivotril)
Plasma – Lithium heparin
40
Phenobarbitone
Gel
100
Phenytoin(Dilantin)
Gel
30
Primidone(Mysoline)
Plasma – Lithium heparin
Valproic acid
Gel
(Epilim)
13
Indications for monitoring anticonvulsants vary between the different drugs.
Suggested guidelines:
•
•
•
•
•
Monitoring initial stabilisation or change of dose–phenytoin, carbamazepine,
phenobarbitone
Suspected toxicity–all drugs
Suspected non–compliance–all drugs
Failure to control seizures–all drugs
Ongoing routine monitoring–phenytoin only, and even this may not be essential.
The therapeutic range is a guide only. Levels below the range may control seizures;
levels above the range may not be toxic and may be necessary for control. When
changing doses, retesting should be delayed a week or so till a new equilibrium
develops.
Phenytoin, carbamazepine and phenobarbitone are potent inducers of hepatic
enzymes thereby raising serum levels of GGT and ALP. The raised enzymes are
regarded as an acceptable side–effect.
Primidone is measured as phenobarbitone which is the active metabolite.
Antidepressant Drugs, tricyclic
Therapeutic monitoring These drugs are not monitored routinely but estimations may
be useful where there is lack of therapeutic response or uncertain compliance.
Drug
Type of Specimen
Anafranil
Plasma – Lithium heparin
Amitriptyline
Plasma – Lithium heparin
Desipramine
Plasma – Lithium heparin
Doxepin
Plasma – Lithium heparin
Imipramine
Plasma – Lithium heparin
Nortriptyline
Plasma – Lithium heparin
Reference ranges supplied with report.
Antidiuretic Hormone (ADH)
Specimen:Plasma – EDTA x 2
Reference Range: Supplied with report
Antimitochondrial Antibodies (AMA)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Antimitochondrial antibodies are seldom found in normal people. Their main use is
in the diagnosis of primary biliary cirrhosis–90% of patients are positive. They are
sometimes present in chronic active hepatitis and occasionally in other autoimmune
disorders.
Anti Mullerian Hormone
Specimen:
Serum – Gel
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Antinuclear Antibodies (ANA)
Specimen:
A
Serum – Gel
Interpretation:
< 1:40
Negative
1:40 or 1:80
Borderline
> 1:160
Consistent with autoimmune disease but not diagnostic
ANA in SLE (Systemic Lupus Erythematosus)
The main indication for ANA is as a screen test in suspected SLE where it will be
positive in 95% of cases. A negative ANA makes SLE unlikely. A positive ANA requires
follow–up by anti–DNA, which has a much higher specificity for SLE, and ENA to
check specificity of the antibody.
ANA in healthy people
The exact incidence of positive ANAs in healthy populations depends on gender
(commoner in women), age–group (rises with age), test methodology, ethnicity etc.
and for this reason precise figures are not available.
A commonly quoted overall incidence is 5%.A recent study narrowed this to 5% in
women aged 40–50. Another quotes 20% in women over the age of 60 rising to 30%
over age 80.
The only certain statement is that positive ANAs are fairly common in healthy people
and should never, as an isolated finding, be represented as possibly indicating SLE
which requires additional laboratory and clinical features before the diagnosis can be
made.
See
Systemic Lupus Erythematosus
ANA in other autoimmune disorders
Incidence of positive ANA in other conditions:
• Rheumatoid arthritis
• Sjogren’s syndrome
• Limited scleroderma (CREST)
• Diffuse scleroderma (systemic sclerosis
• Mixed connective tissue disease (MCTD)
• Polymyositis
• Autoimmune liver disease
30%
60%
50%
80%
80%
30%
< 10%
Drugs causing positive ANA and sometimes a lupus–like illness
• Procainamide–responsible for most drug–related SLE
• Anticonvulsants
• Isoniazid
• Hydralazine
• Chlorpromazine
• Methyldopa.
The clinical manifestations of drug–related SLE are mild and subside within six weeks
of drug withdrawal.
ANA patterns
ANA is detected by adding patient serum to a layer of cells on a slide, then adding a
fluorescent detection system. If ANA is present it shows up as a fluorescent pattern
illuminating various nuclear structures. Usually the pattern is not specific for a
particular disorder but associations have been suggested:
•
•
•
•
Nucleolar pattern
Centromere
Speckled
Peripheral, diffuse
(homogeneous)
•
•
•
•
Diffuse scleroderma
Limited scleroderma, CREST
MCTD, Sjogren’s, polymyositis, RA
SLE.
Their presence indicates vasculitis including:
•
•
•
Wegener’s necrotising granulomatous vasculitis
Systemic vasculitis
Focal necrotising glomerulonephritis with vasculitis.
An initial ANCA screen test has 95% sensitivity for these conditions and is followed by
the more specific antiproteinase 3 (PR3) and myeloperoxidase (MPO) tests.
Antiphospholipid Antibody Syndrome (APS)
A syndrome characterised by widespread, recurrent arterial and venous thromboses
causing ischaemia in the affected organs. Clinical presentations include:
•
•
•
•
Recurrent fetal loss, frequently mid–trimester with placental infarcts
Recurrent arterial or venous thrombosis
Stroke at a young age
Other neurological and cardiological presentations.
APS is found frequently in SLE. When it occurs independently it is known as the
primary antiphospholipid syndrome.
Markers used in the diagnosis of APS are autoantibodies directed against
phospholipids, the main ones being:
•
•
•
Cardiolipin antibodies are used as the initial test when APS is suspected
Lupus anticoagulant
Biological false positive RPR/VDRL.
The aetiology of these antibodies, which may be transient, is unknown but
the presence of elevated titres of anticardiolipin antibodies and/or the lupus
anticoagulants, six months after an episode of venous thromboembolism is a
predictor for an increased risk of recurrence with probable benefit from long–term oral
anticoagulation therapy.
SeeCardiolipin Antibodies
Coagulation Studies
Lupus Anticoagulant/Inhibitor
Capital Pathology Handbook – Interpretation of Laboratory Tests
Antithrombin III (ATIII, antithrombin)
Specimen:Plasma – Sodium Citrate x 2
Reference Ranges:Supplied with report
Antithrombin III is a non–vitamin K dependent physiological inhibitor of coagulation. It
is essential for the anticoagulant effect of heparin.
Deficiency of ATIII, transmitted as an autosomal dominant, is associated with venous
thrombosis. It is a strong but uncommon risk factor, which accounts for < 5% of
patients with thrombophilia. The approximate incidence of the deficiency state
is 1:5,000. Thrombosis occurs in 50% of patients before age 40. It is usually an
indication for life long anticoagulant therapy after a thrombotic event.
ATIII can be spuriously reduced in the presence of heparin and after a thrombotic
event, as well as in nephrotic syndrome, liver disease and pregnancy. Elevated levels
are of no clinical significance.
Apolipoproteins
Specimen:
Serum – Gel
Reference Ranges:Supplied with report
Cholesterol and triglyceride particles in serum are coated with apoproteins to render
them soluble. Quantitation of the various apoproteins provides a more sophisticated
basis for classifying the hyperlipoproteinaemias.
Apolipo–Protein–E Genotyping
Specimen:
Whole blood – EDTA
Reference Ranges:Supplied with report
Apolipo–Protein “a”, A1, B
Specimen:
Serum – Gel
Reference Ranges:Supplied with report
A
APTT (Activated Partial Thromboplastin Time)
Specimen:Plasma – Sodium Citrate
Sodium Citrate tube must be filled to capacity.
Reference Range: 24–37 secs
INVESTIGATION OF COAGULATION DISTURBANCES
Investigation of an increased Prothrombin Time (PT)
Raised-PT
Drugs
Warfarin
• Clinical
• Family history of lifelong bleeding
• Warfarin
No family or life-long
history of bleeding
Factor VII deficiency
(rare)
Mixing test (with normal
plasma)
Correction of
PT
Liver Disease
Vit K. Deficiency
No Correction
Inhibitor
Capital Pathology Handbook – Interpretation of Laboratory Tests
Investigation of Prolonged Activated Partial Thromboplastin Time (APTT)
Increased APTT
Drugs
Heparin
Clinical/family history of
life-long bleeding
No family or life-long
history of bleeding
Factor assays
Mixing test
(with normal plasma)
FVIII, FIX, VWF
Haemophilia A or B
Von Willebrand’s
disease
FIX deficiency
FXII deficiency
Correction
No Correction
Factor Assays
FVIII, FIX,
VWF, FXI,
prekallikrein,
kallikrein, FXII
Lupus inhibitor
Factor specific
anticoagulant
The APTT is a basic test used to detect abnormalities of coagulation involving the
intrinsic pathway. It is used to screen for deficiencies of plasma coagulation factors
(except Factors VII and XIII) or for the presence of an acquired inhibitor. The mixing
test is performed to distinguish between a deficiency and an inhibitor.
The APTT is usually sensitive to factor deficiencies < 40% normal. It is most
commonly prolonged by deficiencies of contact factor FXII, FVIII (reduced in classical
haemophilia and von Willebrand’s disease) and Factor IX (haemophilia B or Christmas
disease).
Marked prolongation of the APTT (> 80 secs) is usually due to haemophilia if
the patient presents with bleeding, or due to Factor XII deficiency if the patient is
asymptomatic.
The lupus anticoagulant, found in the antiphospholipid antibody syndrome, can
prolong the APTT markedly. It is usually associated with an increased thrombotic
tendency rather than bleeding.
Heparin therapy
The APTT is used to monitor standard or unfractionated heparin but not low molecular
weight heparin (LMWH) therapy.
Reference Ranges:APTT ratio: 1.5–2.5
A
Arbovirus
(Ross River, Barmah Forest Dengue Serology)
Specimen:
Serum – Gel
Reference Ranges:Supplied with report
Arsenic
Specimen:Occupational exposure – Random urine
Non-occupational exposure
• 24-hour urine (nil preservative) or random urine or
• Whole blood – EDTA.
Reference Range: Supplied with report
Urine is the preferred specimen for toxicity and occupational monitoring. This method
measures total arsenic. Avoid seafood 5 days prior to collection to exclude non–toxic
organo arsenic compounds.
Arterial blood gases
Specimen:3 mL arterial blood in heparinised syringe transported on ice
Sample should be analysed within 15 minutes (Hospital in-patients).
Includes:
pO2, pCO2, bicarbonate, base excess, pH and oxygen saturation
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Arthritis
For initial investigation (screen), the following tests are recommended:
Antinuclear antibodies (ANA), C- Reactive Protein (CRP), ESR and Rheumatoid
Factor (RF).
Some of the diagnoses to be considered include:
Septic arthritis –immediate joint aspiration (see Synovial aspirate) will give
definitive diagnosis of septic arthritis and differentiate it from gout.
Gout and pseudogout
Rheumatoid arthritis, SLE and other connective tissue diseases
Reactive Arthritis
Reactive arthritis is an aseptic arthritis that develops after an infection elsewhere in
the body. Reiter’s disease, which includes arthritis, conjunctivitis, and urethritis, is an
example of a reactive arthritis. 60–90% of affected patients are positive for HLA-B27
antigen.
• Post–enteric– Yersinia
– Chlamydia
– Campylobacter
– Salmonella
– Brucella
• Post–venereal– Chlamydia
– Ureaplasma
Specific Viral Arthritis
• Arbovirus– Ross River
– Barmah Forest
Arthritis: Infectious
Serology: RRV, BFV, Parvovirus, Rubella, Group A Streptococcus, Yersinia,
Campylobacter, Shigella
Synovial fluid: MCS including cell count and crystal examination
Arthritis: Non-infectious
ANA, Anti-DNA, Anti-ENA, CRP, HLA B27, RF, CCP Ab
ASOT (Antistreptolysin O titre)
Specimen:
Serum – Gel
ASOT
= Antistreptolysin O titre
ADNAse
= AntiDNAse
These tests are used when searching for evidence of recent streptococcal infection in
suspected post–streptococcal glomerulonephritis (PSGN) or rheumatic fever (RF).
Serial tests should be done, looking for rising titres. A high level is more likely to be
significant and sustained levels may indicate persisting infection. PSGN or RF can
easily be over–diagnosed when an isolated, equivocal titre is used as evidence.
See
Rheumatic Fever
A
Aspartate Aminotransferase (AST)
Specimen:
Serum – Gel
Reference Range:Males 3–40 U/L
Females 10–35 U/L
See
Liver function test / interpretation
Aspergillus
A fungus common in soil and decaying material and the cause of an invasive disease
in immunocompromised patients. Diagnosis is by demonstration of hyphae in tissue
biopsy, sputum or culture.
We often recover A. fumigatus and occasionally other Aspergillus species from ear
swabs taken from patients with otitis externa. There is no easily applied antifungal
agent for this situation. Management requires thorough debridement and keeping the
ear dry.
Allergic pulmonary aspergillosis is characterised by asthma and eosinophilia. The total IgE
is often above 1000IU/L. Aspergillus precipitins should be tested for and skin prick testing
arranged.
Aspirin
Specimen:
Plasma – Lithium heparin
Reference Ranges:Supplied with report
Ativan (Lorazepam)
Specimen:
Plasma – Lithium heparin
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Autoantibodies
Specimen:
Serum – Gel
The list of autoantibodies used in the diagnosis of autoimmune disorders continues to
lengthen.
Specificities for particular diseases are seldom absolute and low titre autoantibodies
can be found in apparently disease–free people. Observation over months or years
may show regression, no change or progression to clinical disease.
Each autoantibody is described in more detail under its own alphabetic entry.
Please specify specific tests required according to the patients clinical presentation. If
an ‘Autoantibody profile’ is requested, the following tests will be performed:
Antimitochondrial antibodies (AMA), Smooth muscle antibodies (SMA), Anti Nuclear
Antibodies (ANA), Gastric parietal cell antibodies (GPC) and Thyroid antibodies (TA).
Tissue
Disease
Autoantibody
Adrenal cortex
Addison’s disease
Adrenal
Islet cells
Type I diabetes
GAD 65
IA2
Islet cell (ICA)
Insulin (IAA)
Thyroid
Hypothyroidism
Grave’s disease
Microsomal
Thyroglobulin
TSH receptor
Gastric
Pernicious anaemia
Intrinsic factor
Parietal cell (GPC)
Small intestine
Coeliac disease
Gliadin (AGA)
Tissue Transglutaminase (TTG)
Reticulin
Tissue
Disease
Autoantibody
Liver
Autoimmune hepatitis
1° biliary cirrhosis
Smooth muscle (SMA)
Liver kidney microsomal (LKM)
Mitochondrial (AMA)
MUSK
Neuromuscular
Myasthenia gravis
Acetylcholine receptor
Skeletal muscle
Kidney
Goodpasture’s syndrome
Crescentric glomerulonephritis
Glomerular basement
membrane
Testis
Seminal infertility
Sperm
Skin
Pemphigus
Pemphigus
A
Tissue
Disease
Autoantibody
Haematological
ITP
Post–transfusion
Antiphospholipid (APS)
Platelet
Red cell
Lymphocyte
Cardiolipin
Beta 2 glycoprotein
Connective tissue
disease
Rheumatoid arthritis
SLE
Polydermatomyositis
Scleroderma
Mixed CTD
Sjogren’syndrome
Vasculitis
Rheumatoid factor (RF)
ANA
ds DNA
ENA
CCP
Salivary Duct
Neutrophil cytoplasm (ANCA)
Avian Precipitins (Bird Fanciers’ Disease/Lung)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
B
B
A
Bacterial Swabs
Bacterial specimens are collected for identification of microorganisms and for antibiotic
susceptibility testing. The quality of the specimen is all–important and will determine
whether a pathogen is successfully isolated or whether a false negative culture occurs.
Human pathogenic bacteria like a moist environment, close to body temperature
(37ºC) with no exposure to toxic chemicals. Conversely they are killed by:
•
•
•
Drying
High Temperature
Low Temperature
•
•
•
Sunlight
Antiseptics
Antibiotics.
To provide an optimum environment for swabs, we place them in transport medium
which should be stored at room temperature rather than at 4ºC in the fridge.
Gonococci, for example, survive 24 hours in transport medium at room temperature
but only 2 hours at 4ºC.
An exception is urine swabs which should be stored in the fridge to prevent
overgrowth of contaminants.
Details about collecting swabs from specific sites will be found under their own
alphabetical headings.
•
•
•
•
Cervical
Ear
Eye
Skin
•
•
•
•
Sputum culture
Throat
Urethral
Vaginal.
Bacteroides
An important group of anaerobic gram–negative bacilli. B. ƒragilis, which is part of
the normal flora or the bowel, has the ability to form abscesses when released into
the peritoneal cavity. Specimens from intra–abdominal sites must always be cultured
anaerobically as well as aerobically. Septicaemia and puerperal or post–abortion
sepsis often include B. ƒragilis as a major pathogen.
Bacteroides fragilis produces beta–lactamase which inactivates penicillin and
amoxycillin. Amoxycillian–clavulanate, metronidazole and clindamycin are the most
effective agents.
Bacteroides other than B. ƒragilis are grouped as Bacteroides species. Some may
be sensitive to penicillin. Antibiotic susceptibility testing should be discussed with a
clinical microbiologist.
Some species in the genus have recently been reclassified as Prevotella and
Porphyromonas species.
Barmah Forest Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Basophils
Specimen:
Whole blood – EDTA
Reference Range: Adults < 0.3 x 109/L
The basophil count is part of a routine blood count.
Basophilia is rare as an isolated abnormality. Increases occur in chronic myeloid
leukaemia and other myeloproliferative disorders including polycythaemia rubra vera.
See Blood Count
Bence Jones Protein (BJP)
Specimen:
Random urine
Reference Range: Not present in normal urine
In 1848, Bence Jones described a protein in the urine of myeloma patients that
precipitated at 50ºC and redissolved on boiling. It consists of free light chain fragments
(kappa or lambda) derived from IgG and IgA paraproteins found in serum in myeloma
and occasionally from an IgM lymphoma–associated paraprotein. Because free light
chains are small molecules they pass through the glomerular filter whereas the larger
paraprotein molecules are usually retained in serum.
Presence of BJP in urine is an indication that a paraprotein is malignant rather than
benign. In Bence Jones (light chain) myeloma there is no paraprotein in serum but a
heavy band of free light chains in urine which means that testing for myeloma must
include urine as well as serum electrophoresis.
BJP is detected in two ways:
•
•
As an abnormal band found on EPP (electrophoresis) of concentrated urine
As a kappa or lambda arc on IMF (immunofixation) of concentrated urine
Benzodiazepines (Drug Screen)
Specimen:
Random urine
Reference Range: Not detected
See
Drug Screen
Capital Pathology Handbook – Interpretation of Laboratory Tests
Beta(β) HCG
Specimen:Quantitative HCG
Qualitative pregnancy test
or
Serum – Gel
Serum – Gel
Random urine
Reference Range: Non–pregnant female or male serum : < 5 IU/L
βHCG in normal pregnancy
A single measurement of HCG answers the question whether a pregnancy is present
or not.
Adding a second HCG or a series and plotting them on the HCG graph gives valuable
information on the health of the embryo.
In a normal pregnancy, the HCG doubles every 1–2 days between weeks 3–15
and the trajectory rises parallel to the graph’s percentile lines with remarkably little
deviation.
A plot which is rising parallel to the percentile lines but outside the 5th–95th range
indicates a healthy pregnancy with incorrect dates.
A graph rising more slowly, or flattening, usually indicates the onset of spontaneous
abortion or an ectopic pregnancy and is an indication for proceeding with vaginal
ultrasound looking for an intrauterine gestational sac.
βHCG after spontaneous or induced abortion
After fetal loss or death, the HCG falls over a period of 1–5 weeks depending on its
initial height and the completeness of elimination of placental tissue. The starting HCG
depends on gestational age and health of the conceptus.
βHCG in hydatidiform mole and choriocarcinoma
The abnormal trophoblastic tissue produces large amounts of HCG whose levels
are usually either high normal or clearly above the 95th percentile, sometimes as
high as 1,000,000 IU/L. Ultrasound will separate a hydatidiform mole from a multiple
pregnancy which can also have an HCG level above the 95th percentile.
Absolute HCG levels are more method–dependent in trophoblastic disease and
tumours than in normal pregnancy where levels should be repeatable between
laboratories with different methods. In tumours, there are varying amounts of free beta
chains and beta fragments, and different methods measure these to a varying extent.
For this reason, when monitoring a tumour, specimens should be sent to the same
laboratory.
βHCG in other tumours
Ovarian and testicular germ cell tumours commonly produce HCG and a variety of
malignancies from other sites can occasionally secrete HCG. Levels are used for
monitoring treatment.
B
A
Beta–2–microglobulin
Specimen:Serum – Gel
Urine – random collection.
Reference range: Supplied with report
Beta–2–microglobulin is elevated in autoimmune conditions where there is lymphocyte
activation or destruction.
•
•
•
•
•
•
Multiple myeloma – used for monitoring and prognosis
HIV–levels above 4.0 g/L are more likely to progress to AIDS
Autoimmune disease
Viral infections
Lymphoid neoplasm
Renal impairment.
Bicarbonate
Specimen:
Serum – Gel
Reference range: Adults 20–32 mmol/L
Also known as total CO2, this is a crude acid–base measure. In metabolic acidosis,
bicarbonate is reduced, e.g. in diabetic ketoacidosis. In metabolic alkalosis, it is
increased – as in pyloric stenosis or the compensated respiratory acidosis of chronic
obstructive airways disease.
See flow charts on next page
Capital Pathology Handbook – Interpretation of Laboratory Tests
B
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
Bilirubin
Specimen:Total: Serum – Gel (protect from light)
Conjugated and unconjugated: Serum – clot or gel (protect
from light).
Reference Range:Total:
Adult 4–20 μmol/L
SeeGilbert’s Syndrome
Jaundice
Haemolysis
Hyperbilirubinaemia in infants
I.
Benign Jaundice
•
Physiological jaundice
This is due to breakdown of fetal red cells, “immaturity” of the liver’s glucuronyl
transferase system and increased bilirubin resorption from bowel meconium in infants
as maturation is completed during the first days and weeks of life.
The bilirubin reaches a peak in 5 days (sometimes as high as 300 umol/L) and in
bottle–fed infants disappears in 2 weeks.
•
Jaundice of breast–feeding
This condition, an extension of and inseparable from physiological jaundice, is due to
the presence of maternal hormone in breast milk inhibiting the maturation of glucuronyl
transferase.
Bilirubins can stay high for a long time. In about 2–5%, levels reach 250–350 umol/L
by the 3rd week, remain high for 3–4 weeks, falling to normal over another 1–3
months.
Despite being yellow, the infant thrives, feeds and grows. Changing to bottle–feeding
results in a prompt and sustained fall in bilirubin levels and cessation of breast–
feeding for 24–48 hours, with bilirubins before and after, can be used as a diagnostic
test whilst maintaining lactation by manual expression.
II.
Pathological jaundice
•Haemolytic disease of the newborn
Rhesus incompatibility
The most severe forms of this previously dreaded disease were due to Rhesus
incompatibility between mother, (Rh –ve and with anti–Rh antibodies) and baby
(Rh +ve). The Rhesus system was first described in 1940. Before that time, and until
modern methods of prevention and treatment were developed, these babies were
born dead or anaemic with jaundice at birth increasing rapidly in the first days of life
to cause long–term brain damage in the form of kernicterus. By the late 20th century,
kernicterus had almost entirely disappeared but the fear of it lingers on.
B
A
Haemolytic disease is caused by anti–Rh antibodies in an Rh –ve mother who has
been sensitised during a previous pregnancy with an Rh +ve baby whose red cells
have spilled into the maternal circulation at the time of placental separation. Over the
past 40 years this sensitisation process has been prevented by injecting the mother at
delivery with enough anti–Rh immune globulin to render the fetal cells non–antigenic.
Sensitisation of the mother by an incompatible blood transfusion, unavoidable before
1910, is now an exceptionally rare event.
Despite the success of preventive measures, haemolytic disease is still occasionally
found.
The at–risk mother is Rh –ve and will have red cell autoantibodies when screened
antenatally, usually anti–D, sometimes anti–c or anti–E, least commonly anti–C or
anti–e. For the fetus to be at risk, the father must possess antigen to which antibody is
targeted.
Haemolytic disease in the at–risk infant is diagnosed by examining cord blood at birth,
looking for
•
•
•
Rh +ve infant
Positive Coomb’s test
Hyperbilirubinaemia.
ABO incompatibility and other blood groups
This is more common but rarely severe enough to require exchange transfusion. The
at–risk situation is found in 20% of pregnancies. Usually the mother is Group O and
the infant Group A or B. Prior sensitisation is not required and haemolysis can occur in
the first pregnancy.
Maternal antibody tests are of no use and often the direct Coomb’s test on cord blood
is negative.
Other blood groups include Kell (K), Duffy (Fya, Fyb) and Kidd (Jka, Jkb). For these
incompatibilities, maternal antibodies will be present and the direct Coomb’s on cord
blood positive.
•
Neonatal bacterial infections
Septicaemia is life–threatening and the infant is obviously unwell. Urinary tract
infections are more easily overlooked, the infant being less obviously unwell to begin
with.
•
Hypothyroidism, cystic fibrosis and galactosaemia
Tested on all neonates by way of the Guthrie Card blood spots
•
G–6–PD deficiency
The vitamin K given routinely to neonates elevates bilirubin in G–6–PD hetero–and
homozygotes. On rare occasions homozygous infants (usually males of SE Asian or
Mediterranean origin) develop a severe haemolytic process in the neonatal period,
especially if exposed to certain drugs.
•
Biliary atresia and neonatal hepatitis
The infant is often clinically well during the first few weeks but thereafter there is
an increasing obstructive jaundice shown by rising total bilirubin, a raised level of
conjugated bilirubin, pale faeces and dark urine.
Capital Pathology Handbook – Interpretation of Laboratory Tests
An infant still jaundiced at 3 weeks should have its conjugated bilirubin level
measured–over 20 umol/L of conjugated bilirubin is abnormal and an indication for
further investigation.
When and how often should bilirubins be measured?
As always, clinical judgement and experience are more important than rigid guidelines.
A healthy term baby, growing and feeding well, without visible jaundice, does not need
to have its bilirubin measured.
Premature babies need to be watched more carefully than full–term babies,
particularly during the first 3–5 days.
Jaundice in the first 24 hours of life is not benign. If the jaundiced baby is unwell,
the underlying cause must be identified urgently.
Biopsy Specimens
See
Histopathology, skin biopsy
Blastocystis hominis
A protozoan parasite whose status as a cause of diarrhoea is still unresolved. It can
be observed in the stool of asymptomatic people. Metronidazole, 400–800mg 8-hourly
for 10 days, may be effective treatment.
Blood Banking
Autologous transfusion
Autologous transfusion involves the donation and storage of blood from patients who
will be undergoing elective surgery. If required, these patients can be transfused with
their own blood at surgery. The procedure has become more widespread as concern
over transfusion–transmitted diseases has grown.
1–4 units can be collected pre–operatively at Red Cross House at Woden.
All of these units undergo the same routine screening procedures that are carried out
on all blood collected at the Red Cross. The units are then sent to the laboratory for
storage and crossmatching prior to surgery.
The patient must return to a Collection Centre for a crossmatch specimen to be
collected within five days of the proposed surgery. All of the units that may be
transfused must be crossmatched against this specimen. When required, the
crossmatched units are dispatched to the appropriate hospital or clinic.
Crossmatching requests
Specimen:Whole Blood (EDTA) x 2. Blood should be taken up to 10 days
prior to surgery. If patient is pregnant or has been pregnant in
the last three months or has been transfused in the last 3 months,
blood should be taken no more than 72 hours prior to surgery.
Specimens must be carefully labelled. The following details must be clearly written
on the tube; patients family name, given name, medical record number and/or date
of birth, date and time of collection and signature of the collector.
B
A
Specimens are to be accompanied by a transfusion sheet which must contain the
following details : patients family name, given name, medical record number and/or
date of birth, date and time of collection.
There must be two signatures in the declaration section of the transfusion sheet:
1. The person who collected the blood and;
2.The patient, or if the patient is unable to sign, a witness who can attest to the
patient’s identity.
The declaration should read as follows:
“I certify that I collected the accompanying sample from the above patient whose
identity was confirmed by enquiry and/or examination of their name band and that
I labelled the sample immediately following collection.”
Please provide any relevant clinical information on the transfusion sheet including
previous transfusion, obstetric history and any known antibodies.
The request form and specimen tube will be checked in the laboratory to ensure
validity. Printed sticky labels are acceptable for specimen identification when there
is a signature from the collector on the label.
Emergency transfusion
A pre–transfusion blood sample must be obtained, labelled with the patient’s name,
Medical Record Number, date of birth and date of collection and sent to the laboratory
as quickly as possible. Please contact the Main Laboratory on 02 6285 9811 when
blood is required urgently. A correctly filled out transfusion form is also required.
In an emergency there are units of O negative packed red cells in the theatre fridge’s
for immediate use. The laboratory must be notified immediately if these units are used.
Group and hold (G&H) blood for transfusion
Specimen:
Whole blood (EDTA) x 1
It may be adequate to perform a blood group and antibody screen only (“Group and
Hold”). Blood can then be crossmatched and released promptly if required.
When a Group and Hold has been performed, red cell units can be issued within
30 minutes. Without prior group and hold, a full procedure of blood group, antibody
screen and crossmatch will take approximately 50–60 minutes.
If antibodies are present in the antibody screen, fully compatible units may take even
longer to locate.
Routine crossmatching
Specimen:
Whole blood (EDTA) x 2
Blood should be taken up to 10 days prior to surgery. If patient is pregnant or has been
transfused in the last 3 months, blood should be taken no more than 72 hours prior to
surgery.
Specific blood products
Antihaemophilia Factor (AHF)
AHF is indicated for the management of bleeding episodes and surgical and dental
procedures in patients with haemophilia A (Factor VIII C deficiency) and moderate to
severe von Willebrand’s Disease (vWD).
Capital Pathology Handbook – Interpretation of Laboratory Tests
Cryoprecipitate
B
A
Cryoprecipitate is the cold–insoluble portion of plasma remaining after FFP has
been thawed between 1°C and 6°C. It contains the following factors– fibrinogen,
Factor VIII C, von Willebrand factor, Factor IX, Factor XI, Factor XII and Fibronectin.
Cryoprecipitate is available through the Red Cross Blood Bank, in consultation with
the laboratory.
Fresh Frozen Plasma (FFP)
FFP contains all coagulation factors plus complement and is stored at -25°C in the
laboratory. FFP is indicated for use in control of bleeding in patients with multiple
coagulation defects such as liver disease, dilution coagulopathy and in some
instances of warfarin overdose or if warfarin therapy needs to be reversed.
Platelets
Standard platelet concentrates are available and should be ordered in advance.
For routine purposes at least 24 hours notice is desirable. In an emergency situation
platelets can be provided on request.
The following MSBOS is taken from ANZSBT Guidelines for pretransfusion laboratory
practice 5th edition March 2007 and is intended as a guide only.
Recommended Maximum Surgical Blood Order Schedule (MBOS)
General Surgery
Abdomino-perineal Resection
2
Hiatus Hernia Repair
• Transthoracic2
• Below knee
G&S
• AbdominalG&S
• Above knee
G&S
Incisional Hernia Repair
Amputation
Nil
2
LaparotomyG&S
AppendicectomyNil
LipectomyG&S
Apronectomy/LipectomyG&S
Lumbar Sympathectomy
Bowel Resection
Pancreatectomy2
Anterior Resection
2
G&S
ParotidectomyG&S
Breast Surgery
• LumpectomyG&S
Splenectomy2
• Simple Mastectomy
G&S
ThyroidectomyG&S
• Radical Mastectomy
G&S
Vagotomy and Drainage
G&S
IA*
Varicose Veins Stripping
Nil
Burns Debridement
(*IA = Individual Assessment)
EthmoidectomyNil
CholecystectomyG&S
MastectomyG&S
Colectomy (formation or closure)
G&S
MastoidectomyG&S
Gastrectomy2
RhinoplastyG&S
Gastric Stapling
G&S
TonsillectomyG&S
HaemorrhoidectomyNil
TrachoestomyG&S
Recommended Maximum Surgical Blood Order Schedule (MBOS)
Gynaecological Surgery
Thoracic Surgery
CaesareanG&S
Lobectomy2
ColposuspensionG&S
Pleurectomy2
Cone Biopsy
Nil
Pneumonectomy4
D & C
Nil
Thymectomy2
EctopicG&S
HysterectomyG&S
LaparoscopyNil
MyomectomyG&S
OvariancystectomyG&S
Termination of Pregnancy
Tubil Ligation
Vaginal Repair
G&S
Nil
G&S
VulvectomyG&S
Urological Surgery
Cystoscopy/otomyNil
Cystectomy4
NephrectomyG&S
NephrolithotomyG&S
Prostatectomy
• Open2
• Transurethral resection; TURPG&S
PyelolithotomyG&S
Orthopaedic Surgery
Transurethral Resection of Prostate G&S
ArthroscopyNil
UreterolithotomyG&S
ArthrotomyNil
Femoral Nail Removal
Nil
Fractures
• Femur2
Harrington’s Rods
Hip Replacement
Knee Replacement
4
3
G&S
LaminectomyG&S
MenisectomyNil
Putti-PlattG&S
Spinal Fusion
Synovectomy (Knee)
2
G&S
Vascular Surgery
Aortic Aneurysm – Elective4
Aorto-femoral Bypass Graft
Aorto-iliac Bypass Graft
Av Shunt
4
4
Nil
Carotid EndarterectomyG&S
Femoro-popliteal Bypass Graft
2
Ilio-femoral bypass Graft
4
Sympathectomy Lumbar
G&S
Capital Pathology Handbook – Interpretation of Laboratory Tests
Blood Count (FBC)
Specimen:
Whole blood – EDTA:Adult tube = 4.5 mL blood
: Paediatric tube = 0.5 mL blood
Reference ranges vary widely with age and sex, and detailed reference ranges are
supplied with reports.
Includes Haemoglobin, Haematocrit, Red cell count, Mean cell volume, Mean cell
haemoglobin, Mean cell haemoglobin concentration, White cell count and differential,
platelet count and film examination where indicated.
Blood Cultures
Specimen :
Whole blood taken by aseptic technique
Generally, a blood culture collection requires two culture bottles (aerobic and
anaerobic).
If blood culture for Mycobacteria is required please contact the Microbiology
Department on 02 6285 9846 for special collection details.
Careful aseptic technique is required. The bottles must be labelled (including time
of collection) and if there is any delay in delivery, they should be kept at room
temperature until pick–up.
Collect 10–20 mL into two culture bottles. The volume of blood collected is directly
proportional to the success of isolating an organism. In cases of suspected
endocarditis, three sets over 24 hours are recommend to optimise isolation of the
organism. If antibiotics need to be started rapidly, collect only one set of blood cultures
prior to starting therapy.
Blood cultures are examined continuously and incubated routinely for five days.
Blood Group
Specimen: Whole blood – EDTA
ABO and Rh (D) phenotype are done routinely. The frequencies of the four main
groups of the ABO system in the caucasian population and their naturally occurring
antibodies are:
ABO
Group
Genotype
Frequency (%)
Naturally occurring
antibodies
O
OO
46
anti A, anti B
A
AA/AO
40
anti B
B
BB/BO
9
anti A
AB
AB
5
none
Blood Group and Antibody Screen
Specimen:Whole blood – EDTA
If pregnant please note gestation on the request form.
B
A
Blood Group and Direct Coomb’s Test
Specimen:
Whole blood – EDTA
Blood Group and Hold Serum (G&H)
Specimen:Whole blood – EDTA
Transfusion form and special collection requirements.
See
Blood Banking
Blood Group Phenotyping
Specimen:
Whole blood – EDTA
Bone Marrow
Aspirate and trephine
Bone marrow trephines and aspirates are performed routinely at our main laboratory
at 2 Makin Place, Deakin. Please phone the laboratory on 02 6285 9867 to arrange a
booking. Results will be available as soon as possible.
Patient information sheets available upon request from:
Collection Centres, Capital Pathology website (www.capitalpath.com.au) and from the
main laboratory.
Bone Turnover Markers
See
Deoxypyridinoline Cross Links (DPD)
Hydroxyproline
Capital Pathology Handbook – Interpretation of Laboratory Tests
Bordetella pertussis
Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Antibody (IgA) detection methods are available for diagnosis. For assessing current
illness IgA detection is more reliable later in the disease course. In young children and
infants IgA response may take several weeks to develop, so repeat testing may
be necessary.
PCR
Polymerase Chain Reaction (PCR) requires a plain dry swab with metal wire , which
can be taken as a nasopharyngeal or high oropharyngeal swab. The PCR is highly
sensitive, and is best performed in the early stage of the illness (say the first two
weeks of illness).
Culture
The culture for Bordetella requires a nasopharyngeal or oropharyngeal swab in
transport medium. Culture is not as sensitive as PCR for isolating the causative
organism. PCR is the preferred test in the early disease phase.
Borrelia burgdorferi Antibodies (Lyme Disease)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Brain Natriuretic Peptide (BNP)
Specimen:
Serum – Gel
Reference Range: Supplied with report
BNP is a peptide produced by cardiac atrial cells in response to atrial stretch. The
physiological response to volume overload sensed by atrial stretch receptors is to
induce diuresis by the natiuretic action of BNP on the kidney. BNP can be used in the
assessment of dyspnoea. A normal BNP almost excludes cardiac failure as the likely
cause for shortness of breath, whereas a markedly elevated BNP makes cardiac
failure the most likely cause. Intermediate values for BNP can be found in other
conditions including renal failure, cor pulmonale and pulmonary hypertension.
Breast Aspirate Cytology
Breast Cyst Aspirate – See Cyst Fluids
Solid Lesions – See Fine Needle Aspirate (FNA)
B
A
Bronchial Specimens
Bronchial brushings
This specimen is obtained during bronchoscopy. The brush is placed in a 60mL
specimen container containing saline and vigorously moved around against the
sides of the container to dislodge the adherent cells.
Remove the brush and secure the lid of the container. Specimen should be
refrigerated and transported to the laboratory as soon as possible.
Bronchial washings
Sterile saline (10–20mL) is pipetted down the flexible bronchoscope and sucked
out again into a disposable plastic reservoir.
Specimen should be refrigerated and transported to the laboratory as soon as
possible.
Brucella Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Brucellae are gram–negative bacilli infecting domestic animals, particularly cattle and
pigs. Human infection occurs in farmers and meat–workers as an occupational hazard
or by ingestion of contaminated milk products.
Over the past 30 years, brucellosis has been virtually eliminated from Australia.
Cattle and human brucellosis, when found, will almost invariably have been contracted
abroad.
Agglutination and Coomb’s antibody tests detect both IgM and IgG antibodies. Results
may be negative in the early stages of acute disease and sometimes antibodies
are not detected at any stage. Seroconversion, however, or a > 4–fold increase in
antibody titre, strongly suggests active infection.
Distinguishing previous resolved infection from chronic continuing infection is
impossible using antibody tests alone.
‘A Caring Community’
Providing Maternity, Surgical, Medical and Rehabilitation
services to Canberra and surrounding communities
For a detailed Speciality Directory please email Robbyn Nedeljkovic
[email protected] or 02 6281 8114
www.calvaryjohnjames.com.au
In the Tradition of the Sisters of the Little Company of Mary
with the values of hospitality, healing, stewardship and respect
Capital Pathology Handbook – Interpretation of Laboratory Tests
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C1 (Esterase) Inhibitor
Specimen:Serum – Gel
Specimen should be spun, separated and frozen immediately.
Reference Range: Supplied with report
CA 15–3
Specimen:
Serum – Gel
Reference Range: Supplied with report
Mainly used for monitoring established breast malignancy.
Elevations are found in other benign and malignant conditions.
CA19–9
Specimen:
Serum – Gel
Reference Range: < 40 kU/L
Elevated in pancreatic carcinoma. May be elevated in other gastrointestinal malignancies.
CA 125
Specimen:
Serum – Gel
Reference Range: Age related reference ranges supplied with report
Mainly used for monitoring treatment and progress in established ovarian cancer.
Elevated levels may be found in other malignancies, or occasionally other
non-malignant conditions including endometriosis, Pelvic Inflammatory Disease (PID),
cirrhosis, renal failure and pregnancy.
C-Peptide
Specimen:Serum – Gel
Patient must be fasting.
Reference Range: Supplied with report
Cadmium
Specimen:Whole blood (EDTA)
Or 24-hour urine (nil preservative) or random urine.
Reference Range: Supplied with report
Used where toxicity is a possibility in workers involved with cadmium in industry.
Eating contaminated shellfish may raise levels. If in doubt, repeat after two weeks’
abstention from seafood.
Calcitonin
Specimen:
Serum – Gel. Spin, separate and freeze immediately.
Reference Range: Supplied with report
Calcitonin is a hypocalcaemic hormone produced naturally by the thyroid. There are
no clinical excess or deficiency states.
Calcitonin’s clinical use is as a marker for medullary thyroid carcinoma either as an
isolated tumour or as part of Multiple Endocrine Neoplasia (MEN), type II, along with
phaeochromocytoma and parathyroid tumours.
See MEN (Multiple Endocrine Neoplasia)
Calcium, serum
Specimen:
Serum – Gel
Reference Range: Adults 2.10–2.60 mmol/L
A protein correction is routinely applied:
Adjusted Ca++ = observed Ca++ + {(43-Albumin) x 0.025}
Hypercalcaemia
The two most important causes are:
•
Hyperparathyroidism due to parathyroid adenoma
The parathyroid hormone (PTH) level is increased.
•
Malignant disease
Particularly metastatic tumour in bone, multiple myeloma or cancer of lung, breast
or urinary tract, producing an ectopic PTH–like hormone, PTHrP (PTH related
Peptide).
Other causes of hypercalcaemia include:
• Drugs – Vitamin D
– Thiazides
– Lithium
– Antacids
• Thyrotoxicosis
• Sarcoidosis
• Familial hypocalciuric hypercalcaemia
• Renal transplantation
• Paget’s disease with immobilisation
• Hypocalcaemia
• Chronic renal failure
• Vitamin D deficiency
• Hypoparathyroidism (usually post–thyroidectomy)
• Drugs – Anticonvulsants
– Frusemide
– Biphosphonates
– Oral phosphate
• Malabsorption
• Pancreatitis
• Hypomagnesaemia.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Hypercalcaemia
Hypocalcaemia
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Calcium, urine
Specimen:
24 urine (nil preservative) or random urine
Reference Range: Supplied with report
Some patients with recurrent renal calculi have a high urinary calcium output –
idiopathic hypercalciuria. Serum calcium must be checked to exclude hypercalcaemia.
Campylobacter jejuni/coli
Specimen:Faeces is the usual specimen for acute infection
Serum – Gel for serology
Infections with Campylobacter jejuni or Campylobacter coli cause 60% of bacterial
diarrhoea in the community. These organisms cause an acute enterocolitis which
can be associated with intense abdominal pain. The average incubation period is 3
days; most patients recover within a week. Antibiotic treatment is usually not required
but for severe infections erythromycin is effective. Treatment with quinolones is not
recommended because resistance to them emerges commonly and rapidly. Untreated
patients may excrete Campylobacter in their faeces for 2–3 weeks but transmissibility
is low and it is not usual to put restrictions on otherwise healthy food handlers who are
excreting the organism.
Candida
Candida albicans, the principal pathogenic yeast in humans, is a member of the
normal flora of the gut, respiratory tract and vagina. It can gain dominance under
certain conditions such as diabetes, antibiotic use and suppression of the immune
system. Other species of Candida which are occasionally isolated cause the same
type of infection as C. albicans. For most, the treatment is the same as for C. albicans.
The main sites and types of infection are:
•
Vulvo–vaginitis, predisposing factors are antibiotics, diabetes, pregnancy and
progestagens. Some individuals suffer from recurrent thrush for no detectable
reason. A random glucose should be checked to exclude diabetes.
Specimen: Vaginal swab in transport medium
•
kin, Infection occurs in warm moist areas such as the groin, perianal region,
S
axillae, the breasts or in interdigital webs. It is often seen in those who frequently
immerse their hands in warm water, such as dishwaters.
Specimen: Skin swab in transport medium
•
ails, Candida can cause a painful red swelling of the nail fold resembling
N
pyogenic paronychia. This may progress to nail involvement (onychomycosis).
Specimen:Skin swab in transport medium, nail scrapings for mycology will
grow Candida if present
•
outh, Infections are found mainly in infants and show up as white adherent
M
patches. Laboratory identification is not usually necessary but a swab will grow
the yeast.
•
ystemic candidiasis, Found in immunocompromised patients or in association
S
with prostheses.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Cannabinoids (Drug Screen)
Specimen:
Random urine (nil preservative)
Reference Range: Not detected
Cannabis is rapidly absorbed into fat depots with cannabinoids remaining detectable in
the urine for up to 1–2 weeks after a single exposure. In chronic users, cannabinoids
remain detectable for up to 6 weeks after cessation.
See
Drug Screen
Carbamazepine (Tegretol)
Specimen:Serum – Gel
Trough level should be taken just before next dose (within one hour).
Peak level should be collected 3 hours post dose.
Therapeutic range: 15–40 umol/L
Carboxyhaemoglobin (carbon monoxide)
Specimen:
Whole blood – EDTA
Reference Range: Supplied with report
The affinity of CO for Hb is 200x that of oxygen.
CO toxicity is mainly due to deliberate or accidental exposure to car exhaust fumes.
Levels fall about 15% per hour in air after removal of the CO source. Oxygen
treatment, particularly hyperbaric, accelerates clearance of CO.
Carcinoembryonic Antigen (CEA)
Specimen:
Serum – Gel
Reference Range: Age related reference ranges supplied with report
Cardiac enzymes / markers
SeeTroponin
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Cardiolipin antibodies ( aCL, ACA )
Specimen:
Serum – Gel
Reference Range: Supplied with report
High levels of the IgG antibody are found in:
•
•
•
•
Antiphospholipid syndrome
SLE
Other autoimmune disorders
In some healthy people.
A first indication of aCL may be a “false positive” VDRL or RPR test during routine
antenatal screening.
Low titre antibodies may be transient and are of uncertain significance.
See
Antiphospholipid Antibody Syndrome (APS)
Carotene
Specimen:
Serum – Gel. Protect from light
Reference Range: Supplied with report
Carotene levels are used in the diagnosis of carotenaemia, an orange–yellow
colouration of the skin (but not conjunctivae) that can look like jaundice. The usual
cause is a high intake of vitamin A precursors in carrot or other coloured fruit or
vegetable juice but some systemic illnesses, including hypothyroidism, diabetes,
liver and renal disease, can cause carotenaemia. Because carotene is lipid–soluble,
hyperlipidaemias can give elevated levels. Low values have been used as an indicator
of malabsorption but specificity and sensitivity are poor.
Catecholamines
Specimen:24-hour urine
Plasma special tubes available (Plasma Metanephrine).
Reference Range: Supplied with report
Indications
Phaeochromocytoma
25% of the population have hypertension but only a fraction of these can or should be
screened for phaeochromocytoma. Particular indications are:
•
•
•
•
ymptoms, including sweating attacks, severe headaches, palpitations,
S
nervousness, chest pain; flushing attacks are very uncommon
Episodic hypertension (not always present)
Moderate or severe hypertension in pregnancy or in young people
Adrenal mass.
Neuroblastoma
These usually present as an abdominal mass in children under the age of five.
Number of specimens
When clinical suspicion is low, a single normal result is sufficient but when suspicion is
high, up to three specimens should be tested, preferably collected during or just after
symptoms.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Interpretation
The typical Phaeochromocytoma gives an elevated noradrenaline level. Occasionally
the adrenaline level is elevated but usually it and the dopamine are near normal.
In malignant phaeochromocytomas (10% of the total), dopamine tends to be elevated
as well as noradrenaline.
The typical neuroblastoma shows a huge increase in dopamine.
Non–tumour elevations can be caused by anxiety, stress and exercise, particularly
in the case of dopamine where elevations can be up to twice the upper limit. Small,
isolated elevations of dopamine can usually be ignored.
Essential hypertension can be associated with a minor increase in noradrenaline,
usually less than twice the upper limit.
Medication
Noradrenaline is increased by amphetamines, alpha– and beta–blockers, vasodilators,
theophylline, phenothiazines and tricyclic antidepressants.
Noradrenaline is decreased by clonidine, methyldopa and bromocriptine.
Dopamine is massively increased by levodopa.
Cat Scratch Disease (CSD)
Specimens:
1.Serology
• Paired sera, 2–3 weeks apart.
2. Histology, cytology
• Formalin–fixed tissue, usually lymph node, for histology
• FNA material for cytology.
3. Culture (slow–up to 28 days)
• Whole blood – EDTA
• Tissue or FNA in sterile container.
Cat scratch disease (CSD) is a self–limiting febrile illness with localised tender
lymphadenopathy caused by Bartonella (previously Rochalimaea) henselae which
is transmitted when a bacteraemic cat or kitten bites or scratches a human. In the
United States, where one third of households have a cat, the annual incidence of
CSD is 100 cases/million population, 80% of them in children.
Although CSD was first reported in 1931, it is only recently that B. henselae,
a fastidious slow–growing, gram–negative rod, was identified as the pathogen
responsible. Fleas transfer the organism from cat to cat and flea control is a useful
preventive measure.
Severe systemic illness can occur in immunocompromised individuals.
Treatment
Pain relief may be required. Steroids are generally ineffective. There are no
controlled trials to help choose an antibiotic but for those with prolonged fever and/
or severe lymphadenitis, erythromycin, doxycycline, clarithromycin, ciprofloxacin,
aminoglycosides and sulphamethoxazole/trimethoprim have been used. Therapy
should be discussed with an infectious disease specialist.
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Cerebrospinal Fluid (CSF)
Collect specimen in a sterile plastic CSF tube (These are available from the Stores
Department. Glass tubes cause cellular distortion). Specimen must be received by the
laboratory within one hour of collection due to rapid cellular deterioration.
Please phone a Courier on 02 6285 9877 for an urgent specimen pick up.
Ceruloplasmin (Copper Oxidase)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Ceruloplasmin is reduced in hepatolenticular degeneration (Wilson’s disease), and
Menk’s kinky (steely) hair syndrome.
Oestrogens, anticonvulsants and inflammation cause elevations–ceruloplasmin is a
late acute phase reactant. 95% of serum copper is carried by ceruloplasmin.
SeeCopper
Cervical Cytology
Pap smear
The quality of the Pap smear specimen is of critical importance if false negative
cytology reports are to be minimised.
There are two main reasons for the occurrence of false negative Pap smear reports.
One is laboratory error and the other is sampling error. At Capital Pathology we have
put in place numerous quality assurance measures to minimise laboratory false
negatives. Sampling errors can be reduced by taking an optimal sample. This also
helps the laboratory in the interpretation of the smear.
Specimen collection
Principles of Sampling for Cervical Cytology
The aim of cervical cytology is to detect all cervical pre–cancer. The great majority of
precancerous lesions which we need to detect arise in the transformation zone of the
cervix, an area that is constantly “transforming” during a woman’s reproductive life.
The actual site will vary depending on numerous factors such as hormone influences
or previous treatment. In this area the cells change from glandular to squamous cells,
a process called metaplasia. Metaplastic cells are the transforming cells, which often
have microscopic features of both squamous and glandular epithelium.
The squamous precursor lesions which occur in this area are patchy and they will not
be visible with the naked eye. Glandular precursor lesions often occur in the same
region but may also arise further up the endocervical canal. It is critical then that these
areas are sampled.
Capital Pathology Handbook – Interpretation of Laboratory Tests
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To the clinical observer the transformation zone may appear as a large eversion or
ectropion. It is important to recognise this as a normal process.
Recommended technique for taking a cervical smear
Which instrument?
The appropriate instrument is chosen depending on the appearance of the woman’s
cervix and upon individual experience and preference.
•
The Cervex Sampler (Cervex Brush®) is a broom-like instrument that consists of
numerous specially designed filaments shaped to peak centrally, with shoulders
graded to fit the shape of the cervix.
•
he Cytobrush (cervical brush) is like a miniature bottle brush with small sharp
T
bristles arranged in a spiral around a twisted wire.
•
Spatulas should be plastic and are flat instruments shaped to fit the cervix.
(from top to bottom) Cervex Sampler, cytobrush (cervical brush), plastic spatula
The procedure
Cervex Sampler
Cervex Sampler has a peaked central portion and sloping shoulders. It is applied to
the cervix with the peaked central portion situated in the endocervical canal. Then with
some pressure it is rotated through the full 360° two to three times.
Plastic Spatula / Cytobrush Combination
If choosing a spatula it should optimally be used as a combined technique with a
Cytobrush. The Cytobrush should never be used alone and its use is contra–indicated
in pregnancy. In postmenopausal women and women previously treated for cervical
precancer, the combined spatula and endocervical brush (Cytobrush) method is
appropriate.
Visualise the Cervix
It may be difficult in some patients to visualise the cervix but every attempt needs
to be made to get a clear view of the transformation zone, external os and into the
endocervical canal. The cervix should be visualised under direct illumination with a
speculum in–situ. However, when inserting the speculum it is preferable not to use
lubricant gel as this may cause an unsatisfactory result. Warm water will suffice.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Making The Pap Smear
It is very important to make sure that whatever instrument is used, the sample is
spread evenly and as smoothly as possible along the whole of the glass slide.
Cervex Sampler Technique
Prepare the
conventional smear
by ‘smearing’ first one
side of the brush on the
glass slide, and then
the other. Apply fixative
immediately. If desired a
cervex sampler can then
be used for ThinPrep,
HPV, Chlamydia,
and Gonorrhea.
Spatula Cytobrush Technique
Recommended spatula
cytobrush technique. Most
practitioners prefer to take
the spatula/ectocervical
sample first.
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Fixing the Smear
The smear needs to be fixed as quickly as possible to ensure optimal cell fixation.
Pump spray fixative is recommended. Fixative by immersion in 95% alcohol for 20
minutes is also acceptable. With each technique, the smear needs to be completely
covered. Rapid fixation following smear taking is essential to prevent air–drying, which
can occur within seconds. Sub–optimal fixation is the single most common factor
precluding reliable cytological assessment.
Labelling the Slide
This may be done before or after taking the smear but it is crucial for accurate
patient identification. The slide must be labelled in pencil (ink will wash off during the
normal slide staining and processing) with the patient’s surname, given name and
date of birth. Stick–on labels are unsuitable as these may become detached during
processing.
One Slide or Two
Generally it is preferable to use a technique which makes only one slide. If two
instruments are used, optimal fixation is achieved by smearing the first half of the
slide, covering the unsmeared half with a piece of cardboard and then immediately
spray fixing the smeared half. The cardboard is then removed from the second half of
the glass slide and the second instrument is smeared on this area. The whole smear
can then be spray fixed. Spray fixing the first half again will not affect the final result,
but a delay in spray fixation will be detrimental.
Blood
It is best not to have too much blood on a Pap smear, but sometimes it is unavoidable.
If bleeding should occur during the taking of the smear it is best to proceed, making
sure that squamous and glandular cells are sampled. Smears can be read through a
bloody background but the smear will need to be repeated if it consists only of blood.
The addition of ThinPrep may be worth considering if the smear is bloody.
Mucus
Gently remove any excess mucus from the cervix with a swab. Putting too much
mucus on the slide will result in too scant a sample for accurate diagnosis and will
preclude adequate fixation of the specimen.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Thinprep
ThinPrep is a slide preparation technique that has been shown to be associated with
an increase in diagnostic accuracy of Pap smears.
ThinPrep is a simple procedure, which some studies have shown reduces the risk
of false negative results. The fully automated slide preparation procedure produces
consistent, easy to interpret slides, reducing the incidence of “unsatisfactory” reports,
leading to fewer women being recalled for rescreening.
After preparing a conventional Pap smear, immerse the head of the sampling device
in the ThinPrep vial and agitate the device to release the rest of the cell sample. Do
not break off the head of the device. The ThinPrep vial is then transported to the
laboratory where a ThinPrep slide is prepared. Gentle mechanical dispersion of the
solution frees diagnostically important cells from blood, mucus and cell debris, and a
filtration process results in the collection of epithelial cells which are placed on a slide
in a thin layer prior to staining and microscopic assessment. Lubricant gel may cause
a Thinprep slide to be unsatisfactory.
The resultant ThinPrep slide has a representative sample of cells, relatively free of
mucus, erythrocytes and artefacts.
Some studies suggest that ThinPrep facilitates the identification of more high–grade
lesions than conventional Pap smears.
ThinPrep is available in conjunction with the conventional Pap smear for a cost of $45.
It has no associated Medicare rebate.
When ThinPrep is ordered, according to government regulations, the conventional
Pap smear is not available as a bulk bill test, but can be billed at the rebate level.
If ThinPrep is required, simply endorse the request form by ticking the ThinPrep
Pap test box. The ThinPrep vial can be used for chlamydia and gonorrhoea testing
as well as for HPV testing.
A ThinPrep collection Protocol sheet is available from Client Services Department.
Please discard any expired ThinPrep vials and contact Stores Department for new supplies.
HPV Testing
See Human Papillomavirus (HPV)
Pap Smear Tracking Services
A number of supplementary routine and optional measures are provided to assist
surgeries and their patients. These services may afford valuable and convenient
support for practice follow–up procedures. Please contact the Cytology Department
for any further information.
Tear–off Letters Addressed to Patients
Tear–off letters to the patient, indicating her result and appropriate follow–up, are
attached to the bottom of the Pap smear result report. The patient’s address is printed
on the back of the tear–off section, ready for insertion into a window envelope.These
letters are for doctors to send to their patients if they wish to do so.
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Statistical Analysis of Results
Surgeries are routinely provided with a 12 monthly statistical analysis of the Pap
smears taken. The number and percentage of smears which fall into the negative and
all diagnostic categories are provided. Endocervical cell pick–up rates are also given.
The overall laboratory percentages for all categories are provided for comparison.
A list of patients with abnormal smears in the 12 monthly period is attached to the
statistical report.
Similar information can be provided on request for audit purposes.
Pap Smear Reminder System
Capital Pathology utilises a Pap smear reminder system for all patients with a smear
repeat recommendation of less than two years. The aim of this service, in addition to
other quality assurance measures, is to ensure that the number of women missing
the Pap smear “safety net” is minimised. All letters are sent to the requesting medical
practitioner.
A reminder letter that will be sent to medical practitioners listing all patients with
abnormal or unsatisfactory results requiring repeat Pap smears.
Follow–up letters for patients who have had smears on which colposcopy was
recommended. Doctors are routinely sent questionnaires requesting information on
the diagnostic outcome for patients for whom colposcopy was recommended. (As
part of our quality assurance protocols, we are required by government regulations,
to correlate all high grade or inconclusive Pap smear results with any further clinical
information.)
Additional Pap Smear data lists are available as required. Please contact Cytology
Department to discuss further on 02 6285 9867.
Capital Pathology has been working closely with the ACT Cervical Cytology Register
since its inception and is involved in its management and advisory committee
functions. Unless the patient has opted off the Pap test register, her Pap test results
will be automatically sent to the Pap Test Register (as required by legislation).
Capital Pathology also works closely with the NSW Pap Test Register.
Cervical Swabs
See
Chlamydia trachomatis
Neisseria gonorrhoea
Capital Pathology Handbook – Interpretation of Laboratory Tests
Chlamydia pneumoniae
Specimen:Serum – Gel
Preferably paired sera.
Reference Range: Supplied with report
C. pneumoniae is recognised as an important cause of atypical pneumonia, second
only to Mycoplasma. Like the other Chlamydia it is an obligate intracellular bacterium.
Spectrum of disease
Many adults have antibodies indicating that the infection is common, initial infection
being typically at age 5–15 years. It can cause pneumonia, severe pharyngitis,
hoarseness, fever, cervical lymphadenopathy. Infection in young adults is usually
of mild to moderate severity but can be sub–clinical or, in immunocompromised
patients, severe. Incubation period averages 21 days and infection can recur. Its most
recent (and surprising) association is with coronary artery disease, suggested by
seroepidemiologic studies and finding the organism in atheromatous plaques.
Laboratory diagnosis is typically based on serology.
Chlamydia psittaci
Specimen:Serum – Gel
Preferably paired sera.
Reference Range: Supplied with report
C. psittaci is a pathogen endemic in all bird species. When a human inhales dust
from fomites from infected birds, they can develop an infection which may present as
an atypical pneumonia, headache, fever, rash, myalgia. Severity ranges from mild to
moderate, occasionally severe.
Psittacosis is largely confined to bird–fanciers (the parrot family particularly) and
poultry–handlers.
Laboratory diagnosis is based on antibody findings.
Chlamydia trachomatis
PCR (Polymerase Chain Reaction) testing is the diagnostic technique routinely used
by Capital Pathology to detect Chlamydia trachomatis.
This procedure uses nucleic acid probes which are specific for all serovars of C.
trachomatis. The main advantage of this technique is through the step of PCR target
amplification. Here, specific DNA sequences of the cryptic plasmid are amplified
exponentially, making their presence easy to detect by a colour formation step.
Enzyme Immuno–assay (EIA) and the Direct Fluorescent Antibody (DFA) techniques are
no longer used as they have been found to be less sensitive and less specific. The PCR
technique detects both “live” and “dead” Chlamydia (as do DFA and EIA), but experience
indicates that treated infections will test as “negative” four weeks after treatment.
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Specimen requirements
As with all laboratory testing, the accuracy of the result is affected by the quality of the
specimen.
Chlamydia are Gram–negative, non–motile organisms that, due to their inability
to synthetise ATP, exist as obligate intracellular pathogens of columnar, and not
squamous epithelium. Appropriate cellular material must be collected in order to detect
the organism.
Urine Collection
Urine specimens offer advantages in terms of ease of collection and patient comfort.
PCR does not necessarily require cervical or urethral swabs, with the discomfort
and inconvenience inherent in these procedures. It allows a prompt diagnosis of
genitourinary chlamydia infection, in both males and females, using the first 20–30mL
of the stream.
Patients should not have urinated for the previous two hours. Urine PCR replaces
urethral swabs for diagnosis in all males and females in whom visualisation of the
cervix is not indicated.
Cervical Samples
Remove mucus from exocervix with a large swab and discard. Insert another large
swab into endocervical canal until tip is no longer visible. Rotate 3–5 seconds.
Withdraw. Avoid contact with vaginal surfaces. If patient is also having a ThinPrep
pap smear, then Chlamydia PCR can be done on the ThinPrep fluid. Please request
“Chlamydia PCR testing” on request form.
Specimen Transport – Female Genital samples
Directly after sampling, vigorously agitate swab in Cobas Specimen Transport Medium
for 15 seconds. Express liquid against side of the tube. Excess mucus should be
removed by collecting it on the swab. Express any excess liquid from the mucus
against the side of the tube. Remove swab and any excess mucus and discard.
Specimens should be stored at 4°C and transported to the laboratory as soon as
possible.
Chloride
Specimen:
Serum – Gel
Reference Range: 95–110 mmol/L
Capital Pathology Handbook – Interpretation of Laboratory Tests
Chlorpromazine (Largactil)
Specimen:Serum – Plain clot
Do not use gel tube.
Collect pre dose (trough) specimen just before next dose.
Reference Range: Supplied with report
Cholesterol
Specimen:
Serum – Gel
Total cholesterol is made up of three fractions:
Total = LDL + HDL + VLDL/IDL
Total and HDL cholesterols are measured directly in the laboratory
VLDL/IDL is estimated from the fasting triglyceride using the formula:
Triglyceride
2.2
LDL is calculated by subtraction using the Friedewald formula:
VLDL=
Triglyceride
2.2
The formula is inaccurate and not used when the triglyceride is above 4.5.
LDL=total–HDL–
Desirable range: the aim of lipid lowering therapy is to obtain a:
Total cholesterol of < 4.0
LDL cholesterol of < 2.5
HDL cholesterol of > 1.0
Triglycerides of < 2.0
See
Lipid Disorders
Triglyceride
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Cholinesterase
Specimen:•Whole blood (Lithium Heparin) is preferred because it
can be used for both red cell and plasma cholinesterase
measurements when checking for insecticide poisoning
and can be used in detecting scoline sensitivity
• Serum can be used, however, please specify serum
cholinesterase is required
•Please include clinical details.
Indications:
1. Chronic exposure to organophosphate and carbamate anticholesterase sprays.
Organophosphates
Carbamate
Malathion
Carbaryl
Acephate
Methiocarb
Coumaphos
Methomyl
Chlopyrifos
Propoxur
These insecticide sprays are widely used by horticulturists.
Red cell and plasma cholinesterases should be measured before spraying
commences to establish the individual’s baseline reference range and at regular
intervals thereafter.
If the baseline is unknown, estimations at 3–day intervals after removal from exposure
will show recovery towards the baseline.
Because red cell cholinesterases are irreversibly inhibited by organophosphates
(but not carbonates), levels remain low for the 4 month life of erythrocytes. The red
cell level is the preferred test for monitoring low level chronic exposure.
2. Acute poisoning
Plasma cholinesterases fall sharply when acutely exposed to organophosphates
or carbamates but recover to their previous levels within less than a week.
Measure plasma (or serum) cholinesterase.
3. Scoline (suxamethonium) sensitivity
Measure serum or plasma cholinesterase and dibucaine number. Do not test within
one week of scoline administration or two weeks of blood transfusion.
4.Other
Cholinesterase levels are reduced in chronic liver disease, renal disease, pregnancy/
oestrogens, acute illness.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Chronic Lymphocytic Leukaemia (CLL)
Typically CLL is an indolent disorder with a median survival of 10–14 years. In
the early stages the patient does not require therapy but regular blood counts are
performed to monitor the disease for increased activity.
Diagnosis is based on peripheral blood finding of lymphocytosis with cell marker
analysis to demonstrate a monoclonal B cell population with light chain restriction.
Bone marrow biopsy is not essential for diagnosis but is useful to determine bone
marrow reserve and possibly outcome prediction.
The indications for treatment are:
•
•
•
•
Bone marrow failure with anaemia, neutropenia or thombocytopenia
Bulk disease with lymphadenopthy and/or hepatospenomegaly
Constitutional symptoms such as fever, weight loss, night sweats
Autoimmune complications, usually haemolytic anaemia.
Predictors of a poor outcome:
1. Clinical stage, RAI or Binet staging system
• > 3 nodes with bone marrow failure: mean survival 2 years
• > 3 nodes: mean survival 7–8 years
• < 3 nodes with normal haemoglobin and platelets: approaches age–matched
control survival.
2. Lymphocyte doubling time < 12 months
3. Abnormal karyotype on chromosome analyses
4. Marrow infiltration pattern, nodular being better than diffuse.
Treatment is not given on the basis of the high lymphocyte count alone. Initial
therapy usually involves the alkylating agent chlorambucil with or without prednisone.
Fludarabine is one of a new group of agents which is an alternative first line treatment
for CLL.
Chronic Myeloid Leukaemia (CML)
CML is predominantly a disorder of middle life but the diagnosis is being made
increasingly in younger patients. The disorder is characterised by a leucocytosis which
is left shifted, typically showing the major increases in myelocytes and segmented
neutrophils giving a bimodal differential white cell distribution. Blast cells are also
present. The majority of patients are Philadelphia chromosome positive. The NAP
score is low or absent.
CML is a state of bone marrow instability and after a variable time, median 3–4 years,
the disease transforms into an acute leukaemia which is unresponsive to standard
chemotherapy. The only potentially curative treatment is bone marrow transplantation.
This may use a matched sibling donor or a matched unrelated donor transplant.
Interferon is the treatment of choice in patients who cannot undergo bone marrow
transplantation. Hydroxyurea is frequently used for cytoreduction.
C
A
Chyluria
Specimen:
Random urine, ask for triglyceride
Chyluria is due to blockage of lymphatics, usually by filaria but sometimes by
malignant disease. The urine is milky due to triglyceride–rich chylomicrons in the
lymph.
CK / CKMB
SeeTroponin
Clonazepam (Rivotril)
Specimen:Plasma – Lithium heparin
Trough level is suggested, taken just before the next dose
(within one hour).
Reference Range: Supplied with report
Clostridium difficile
Specimen:
Faeces for C.difficle toxin
Reference Range: Not detected
A normal inhabitant of the bowel which can proliferate when other organisms are
suppressed by antibiotics, particularly clindamycin or ampicillin. C. difficile produces
toxins causing bloody diarrhoea and pseudomembranous colitis visible on endoscopy.
Less severe forms of antibiotic–associated diarrhoea may also be associated with C.
difficile.
Clostridium Tetanus Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Clozapine
Specimen:Whole blood – EDTA
Trough level at least 6 hours post dose. If peak level requested,
collect at one hour.
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Coagulation Studies
Specimen:Plasma – Sodium citrate and EDTA
Sodium citrate tube must be filled to capacity.
Please request specific testing depending on clinical indications.
Anticoagulant Therapy
Heparin
The routine test for monitoring heparin therapy is the Activated Partial Thromboplastin
Time (APTT).
Specimen:
Plasma – Sodium citrate
Therapeutic range: 45–79 seconds
Ratio:1.5–2.5
Notes:
• It is most important to obtain the correct volume of blood (i.e. exactly 4.5 mL
added to the tube).
• The blood must not be collected from a limb being used for intravenous infusion.
• A periodic platelet count is recommended when monitoring heparin therapy
Specimen required is 1 x 5 mL whole blood (EDTA).
Warfarin Therapy
The routine test for monitoring warfarin therapy is the International Normalised Ratio
(INR).
Specimen:Plasma – Sodium citrate
The blood must be tested within six hours of collection
and should be kept at 4–6 °C.
Therapeutic range:2.0–4.5
Coagulation Screen
The following coagulation screen, in conjunction with a clinical history, provides the
basis for the assessment of patients with suspected coagulation disorders.
The coagulation screen includes:
1.FBC
2. Prothrombin Time (PT)
3. Activated Partial Thromboplastin Time (APTT).
4. Thrombin Time (TT)
5.Fibrinogen.
Specimens: 1 x EDTA
1 x Sodium citrate
Reference Range:PT
10–13 seconds
APTT 24–37 seconds
TT
10–15 seconds
C
A
Notes:
• It is most important to obtain the correct volume of blood (4.5 mL of blood added
to tube) in the citrate tube.
• Traumatic venepuncture and/or long delay between collection and courier pick–up
should be avoided, as should short collections. These may all affect the result.
• Skin bleeding time is not included as part of a routine screen.
D–Dimer
This screening test is elevated in patients with disseminated intravascular coagulation
(DIC), pulmonary embolism (PE) and deep vein thrombosis (DVT).
Raised levels as an indication of reactive fibrinolysis have been reported in: Sickle Cell
Disease, Liver Disease, Severe infection / sepsis, Pre–eclampsia and Malignancy.
Specimen:1 x Sodium citrate
Factor Assays
Specimen: 2 x Sodium citrate
The specimens must be separated and frozen as soon as possible and transported on ice.
The Importance of the Bleeding History
A careful history is the most valuable part of an assessment. If the history of bleeding
is convincing, a minor disorder may exist even if the laboratory tests are normal.
Ask specifically about:
•
•
•
•
•
•
•
•
EpistaxisEspecially if bilateral, recurrent, or requiring repeated cautery–
unilateral epistaxis is often due to a local lesion.
BruisingSpecify size and whether spontaneous or traumatic; trunk and
upper arm bruising are potentially more important than lower limb
bruising–deep bruising and central induration is significant.
Lacerations Particularly recurrent and prolonged bleeding
SurgeryWisdom tooth extraction, tonsillectomy, other major surgery, blood
transfusion requirement, prolonged postoperative hospital stay.
Menorrhagia A bleeding disorder may potentiate anovulatory menhorragia.
Gum bleeding When brushing teeth
Family history Suggests inherited disorder
DrugsSpecifically ask about medication for headaches, arthralgias,
particularly aspirin, NSAIDs.
Cocaine (Drug Screen)
Specimen:
Random urine: (nil preservative)
Reference Range: Not detected
See
Drug Screen
Coeliac Disease (Gluten Sensitive Enteropathy)
Coeliac disease, affecting at least 1:3000 (1:300 in some populations) is by far the
commonest cause of intestinal malabsorption.
In severe cases it presents in infancy or childhood with failure to thrive and fatty
diarrhoea.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Milder cases are more common and can easily be overlooked, particularly as many do
not have intestinal symptoms.
Features of these milder cases can be one or more of:
•
•
•
•
•
Anaemia
Iron deficiency
Folate deficiency
Fatigue
Diarrhoea, abdominal discomfort.
The disease is due to an intolerance to gliadin (gluten) found in wheat, barley and
rye. Diagnostic antibodies are found in serum, and the mucosa of the small intestine
shows flattened villi on biopsy. Withdrawal of gluten from the diet usually results in
symptomatic improvement, normalisation of villi and disappearance of antibodies over
the course of a year or so.
Dermatitis herpetiformis, which presents as a chronic pruritic papulovesicular skin
rash, is a cutaneous manifestation of gluten sensitivity with the same diagnostic
antibodies and improvement on gluten withdrawal.
When to test for Coeliac Disease
It is recommended that doctors test for the disease in patients with:
•
•
•
•
•
•
•
•
•
•
GI symptoms (eg bloating, abdominal pain)
Failure to thrive
Type 1 DM
Turner syndrome
Down syndrome
Family members who have coeliac disease
Iron-deficiency anaemia
Unexplained liver function test abnormalities and, in children, cryptogenic cirrhosis
Osteoporosis
Malabsorption syndromes.
Many disease states have been linked with coeliac disease and more commonly,
gliadin antibodies. These include inflammatory arthritis, peripheral neuropathy, ataxia
and infertility (including miscarriages). The evidence behind these associations is
often weak, and in the case of gluten-sensitive ataxia, recent studies have essentially
debunked the hypothesis. Infertility remains an area of uncertainty, though GESA
suggest testing may be appropriate.
Testing recommendations
• Serum IgA TTG (order through TTG) and serum IgA.
• If IgA deficient, order IgG gliadin and consider referral to a gastroenterologist or
clinical immunologist if clinically indicated.
• In high risk individuals (i.e. with associated conditions) order IgA TTG and if
negative, consider ordering coeliac tissue typing.
• A negative DQ2 and/or DQ8 result significantly reduces the risk of having or
developing coeliac disease (less than 1% in DQ2/8 negative patients)
• A positive DQ2 and/or DQ8 status confers an absolute risk of up to 4% of having
or developing coeliac disease, and should not in itself be used to diagnose or
label a person as being at high risk of having or developing coeliac disease.
C
A
Cold agglutinins (Cold Antibody Titre)
Specimen:Serum – Plain clot
Specimen to be kept at 37ºC until processed.
Reference Range: Not detected
Complement, C1q
Specimen:
Serum – Gel
Reference Range: Supplied with report
Complement Fractions C3 and C4
Specimen:
Serum – Gel
Reference Range: Supplied with report
The complement system is a cascading series of plasma proteins whose end–
products cause bacterial lysis and remove the immune complexes found in post–
streptococcal glomerulonephritis, SLE and other autoimmune disease. There is
reduction of C3 in PSGN and of both C3 and C4 in SLE. They are increased in most
other inflammatory states.
Complement, Total Haemolytic (CH50, CH100, Total Haemolytic Complement)
Specimen:Serum – Gel
Serum should be spun, separated and frozen within 20 minutes
of collection.
Reference Range: Supplied with report
Congenital Adrenal Hyperplasia (CAH)
A group of inherited adrenal disorders due to enzyme defects, the commonest being
21–hydroxlylase deficiency by measuring 17–OH progesterone.
Severe CAH can present as hypokalaemia and dehydration in neonates; as virilisation
in a female child; or as precocious puberty in males.
A milder and more common form of CAH can present as hirsutism in adult women. It
can be detected by measuring serum 17–OH progesterone on a morning specimen
collected during the follicular phase of the menstrual cycle.
See
17–Hydroxy Progesterone (17-OHP)
Capital Pathology Handbook – Interpretation of Laboratory Tests
Connective Tissue Diseases
A group of systemic autoimmune diseases characterised by the presence of
fairly non–specific autoantibodies and associated with chronic inflammation of
musculoskeletal structures. The main diseases are:
•
•
•
•
•
•
•
Rheumatoid arthritis (1–2% of the population)
SLE, Systemic Lupus Erythematosus (0.1% of the population)
Sjogren’s syndrome
Diffuse scleroderma (0.002% of the population)
Local scleroderma (CREST)
Mixed connective tissue disease (MCTD)
Dermato/poly/myositis.
The following tests may be useful : Anti Nuclear antibodies (ANA), Double stranded
DNA (DS DNA), Rheumatoid factor, and Extractable Nuclear Antigens (ENA).
Diagnosis can be difficult or impossible in the early stages of these conditions which
can evolve over months or years before enough features develop to establish the
diagnosis.
Coomb’s Test
Specimen:
Whole blood – EDTA
Reference Range: Supplied with report
The Coomb’s test detects antibodies directed against red cells.
The direct Coomb’s test detects antibodies or complement which are coated on red
cells as in autoimmune haemolytic anaemia, haemolytic disease of the newborn,
incompatible transfusions, or drug–induced haemolysis, particularly that due to
methyldopa or penicillin in large doses.
The indirect Coomb’s test detects red cell antibodies in serum, as in maternal antibody
screens in pregnancy or some autoimmune haemolytic anaemias. It is also used in
cross–matching.
C
A
Copper
Specimen:
erum – Gel
S
or
Urine, 24-hour (nil preservative).
or
Liver biopsy placed in sterile container without formalin or saline
Occupational exposure – Urine.
Reference Range: Supplied with report
Wilson’s disease is an autosomal recessive disease due to accumulation of copper
in the body in toxic amounts. It presents usually at age 5–20 with unexplained liver
disease, neurological or psychiatric symptoms, or Kayser–Fleischer corneal rings.
There is an increased excretion of urine copper, but reduced serum copper. This is
because ceruloplasmin, the protein which transports copper in serum, is reduced in
Wilson’s disease even though the total body load of copper, and its urinary excretion,
are markedly elevated.
S. copper is reduced in protein–losing states and raised by oestrogens, pregnancy, or
inflammatory states.
Cord Blood Testing
When haemolytic disease of the newborn is suspected, it is recommended that the
cord blood is tested for Haemoglobin, Blood Group and Direct Coombs. If the direct
Coombs is positive, further typing is performed and the coating antibody identified
Cortisol, serum
Specimen:
Serum – Gel
Reference Range:a.m. 100–535 nmol/L
p.m. 80–480 nmol/L
There is marked diurnal variation, the peak at 09.00 hours being 50–100% higher than
the trough at 23.00 hours.
Decreased levels
• Primary adrenal insufficiency (Addison’s disease)
• Secondary deficiency follows adrenal suppression by steroid therapy
• Drugs–ketoconazole, phenytoin, metyrapone, steroids
• Decreased cortisol–binding proteins
• Hypopituitarism
• Exogenous steroids (variable).
Elevated levels
• Cushing’s syndrome
– pituitary adenoma or hyperplasia
– adrenocortical tumour
– ectopic ACTH from malignant tumour
• Stress, illness, depression, alcoholism
• Oral contraceptives, oestrogens, pregnancy
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
•
Exogenous steroids (variable)
Increased cortisol–binding proteins.
Cortisols are of limited value when monitoring replacement therapy.
SeeCushing’s Syndrome
Cortisol, urine
Specimen:
24 hour urine (nil preservative)
Reference Range: Supplied with report
Because only the unbound fraction of serum cortisol reaches the urine, this is a good
screen test for Cushing’s syndrome and a clearly normal result makes the diagnosis
unlikely. In perhaps 10% of cases cortisol hypersecretion is intermittent.
Elevations up to 3x normal may be found in stress, depression or alcoholism.
Corynebacterium diphtheriae
A special medium is required to recover C. diphtheriae so the possibility of diphtheria
must be specifically mentioned on the request form.
Tonsillar diphtheria is still occasionally seen in non–immunised persons. Skin
infections with C. diphtheriae occur in the tropics and can be a source of infection.
Coxsackie Viruses
These are widely distributed enteroviruses associated with many different types of
illness including minor febrile illnesses, the common cold, herpangina, pleurodynia,
aseptic meningitis, myocarditis, post–viral fatigue syndrome, conjunctivitis, Type 1
diabetes and others. Infections are more common in summer and autumn. Virus can
be recovered from throat swabs or rectal swabs and also conjunctival or vesicular
swabs if lesions are present.
C–Reactive Protein (CRP)
Specimen:
Serum – Gel
Reference Range: < 5 mg/L
CRP is the most useful of the acute phase reactants, rising sharply 4–8 hours after
tissue damage by infection, inflammation or trauma. It returns to normal 2–3 days after
disease activity has ceased. It can be regarded as a fast–changing ESR which, by
contrast, rises and falls more slowly. It can be used as an indication of occult bacterial
infection, suspected rheumatic fever, inflammatory bowel disease or other conditions
where there is uncertainty whether symptoms are functional or due to organic disease.
Chronic inflammatory diseases such as SLE can be monitored using serial CRPs and
for this purpose it is a more useful test than the ESR.
C
A
Creatine Kinase (CK)
Specimen :
Serum – Gel
Reference Range:Males 5–190 U/L
Females 5–165 U/L
Causes of an elevated CK – Possible causes:
Cardiac muscle
• Infarction
• Myopathy
• Myocarditis.
Miscellaneous
• Macro–CK
• Malignancy
• Cerebrovascular disease
• Diabetic ketoacidosis.
Skeletal muscle
Useful further tests
Troponin
Serum LD and AST
CK–Isoenzymes
Urinary myoglobin
Autoimmune immunology
DNA tests for muscular dystrophy
Injury/Trauma
Crush; Surgery; IM injections;
Ischaemia
Alcohol
Acute/Chronic alcohol excess
Infection
Influenza; Coxsackie A and
B; Clostridia; Streptococcus
pyogenes; Parasitic infestations
Endocrine
Hypothyroidism;
Hyperthyroidism; Steroid
myopathy
Metabolic
Hypokalaemia; Vitamin D
deficiency; Carnitine deficiency;
Carnitine Palmitoyl–tranferase
deficiency; Hypoparathyroidism
Autoimmune
Polymyositis; Dermatomyositis
Exercise
Severe exertion; Marathon
run; Convulsions; Paroxysmal
myoglobinuria
Heat stroke
Malignant hyperpyrexia
Muscular dystrophy
Developed by N. Walmsley 1995. Adapted with Permission.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Creatinine, serum
Specimen:
Serum – Gel
Reference Ranges:
age
µmol/L
up to 7 days
20–60
8 days–4 years
20–40
5 years–11 years
30–70
Adult male
60–110
Adult female
40–80
Serum creatinine is widely used as a test of renal function both as a general screen,
along with urine protein, for renal disease, and as a test for serial monitoring of renal
function.
Creatinine reflects glomerular filtration rate (GFR), and it also reflects endogenous
creatinine formation from skeletal muscle and to a much lesser extent, exogenous
creatinine excretion from cooked meat.
GFR steadily reduces with increasing age but this is offset, though not completely, by
diminishing muscle bulk and creatinine excretion.
The use of population–based reference range is even less satisfactory for creatinine
than for other analytes because the individual creatinine range for a person remaining
in good health is narrower than the traditional population range. When monitoring a
potentially nephrotoxic process, reference should always be made to the individual’s
own range, as shown by creatinine results when disease–free, rather than to the
population range.
Uses of serum creatinine
• To estimate GFR according to the MDRD formula (reference: Position Statement,
The Australian Creatinine Consensus Working Group. Medical Journal of
Australia, Vol. 183:3 pp138–142, 2005)
• For establishing an individual’s baseline creatinine range
• Monitoring potentially nephrotoxic drugs, particularly in the elderly
–NSAIDs
– ACE inhibitors
–Aminoglycosides
–Diuretics
–Lithium
–Others
• Monitoring potential nephropathy e.g. in diabetics
• Monitoring established renal disease
• Monitoring renal transplant rejection
• Monitoring renal dialysis.
C
A
Creatinine, urine
Specimen:
24 hour urine, nil preservative
Reference Range: Supplied with report
24 hour urine creatinines are used when estimating creatinine clearance.
Wide variations in creatinine output for an individual are due to biological variation
of ±20%. Sometimes compounded by incomplete (or over–complete) urine collects.
A creatinine concentration in a spot urine gives a way of compensating for urine
concentration when expressed as the ratio, analyte/creatinine e.g. ACR (albumin/
creatinine ratio) in diabetics.
Creatinine Clearance
Specimen:24 hour urine (nil preservative) and serum gel
Must be accurately timed to 24 hours, plus serum obtained
within collection period. Please report height and weight on
the specimen.
Reference Range: 70–150 mL/min
CREST Syndrome
•
•
•
•
•
Calcinosis
Raynaud’s phenomenon
Esophageal hypomotility
Sclerodactyly
Telangiectases.
Also called limited scleroderma.
The ANA is usually positive showing the centromere pattern.
Creutzfeldt Jacob Disease
This is a prion disease where the clinical manifestations result from an accumulation
of an altered prion protein molecule in the central nervous system. The diagnosis is
suspected from the history and clinical examination. Confirmation of a case can be
made by histology on brain biopsy or post mortem tissue supported by molecular
biology techniques to look for expression of gene sequences.
Please contact the Director of Clinical Pathology to discuss further details on 02 6285 9895.
Crohn’s Disease
Laboratory abnormalities can include:
•
•
•
•
•
•
Iron deficiency
Vitamin B12 deficiency
Anaemia due to combinations of chronic disease and iron and B12 deficiency
Raised ESR and CRP
Reduced albumin
Hypokalaemia.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Cryoglobulin and Cryofibrinogen
Specimen:Cryoglobulins – Serum (plain clot)
Cryofibrinogen – Plasma (Sodium Citrate) and plain clot.
Reference Range: Not detected
Specimen must be kept at 37°C prior to analysis
See
Cold Agglutinins (Cold Antibody Titre)
Cryptosporidium, faeces
Cryptosporidium is now recognised as a cause of acute gastroenteritis, particularly
in children. It is a notifiable disease. It is found in a variety of hosts and transmission
from farm livestock or pets to humans can occur. Person to person transmission also
occurs and has been responsible for outbreaks in child care facilities.
Diagnosis is by use of a special stain for oocysts in faeces and will be done on specific
request. The typically watery diarrhoea usually settles without treatment within 10
days (range 1–20 days). In immunocompromised patients (especially those with HIV),
it may cause a severe prolonged diarrhoeal illness. In this situation specialist advice
should be sought on treatment.
Cryptococcal Antigen
Specimen:
Serum – Gel or Cerebrospinal Fluid
Reference Range: Not detected
Crystal Identification
SeeHistopathology
Synovial Aspirate
Cushing’s Syndrome
The main causes are:
• Excess ACTH (adrenocorticotrophic hormone) produced by the pituitary
• Excess ACTH produced by non–endocrine tumour particularly lung
• Adrenal tumours.
Basic screen test: 24-hour urine free cortisol
Follow–up test:
Dexamethasone suppression test
Isolated serum cortisol is not recommended as a screen test though a level below
500 nmol/L in a specimen collected before 10.00 hours makes Cushing’s unlikely.
Clinical features of Cushing’s include obesity, diabetes, hypertension, plethora, muscle
weakness, striae and osteoporosis.
Cyclosporin
Specimen:
Whole blood – EDTA
Reference Range: Supplied with report
C
A
Cyst Fluids
Cyst fluids should be placed into a labelled specimen container and refrigerated at
4ºC. Storage overnight is satisfactory. It is preferable to send the whole specimen to
allow for concentration of poorly cellular specimens and for preparation of a cell block.
The cell block is then available for special staining including immunohistochemistry
should this be necessary for the diagnosis.
If the volume of cyst aspirate is very small (i.e. several drops only), smears can be
made directly from the fluid. These should be rapidly fixed with pump spray fixative.
For greater detail on smear preparation please see section on Fine Needle Aspirate
(FNA). Alternatively a small amount of normal saline can be added to the specimen
container. Please note the volume of saline added on the request form.
Cystic Fibrosis PCR
Specimen:
Whole blood – EDTA x 2
Two generations of family tree needed with details of any cases of cystic fibrosis.
Need ethnic background, e.g. Caucasian etc.
Affects 1:3000 infants. Cystic fibrosis is carried on a recessive gene which can be
identified in 70% of carriers by DNA testing. The underlying defects of exocrine
gland function show up in respiratory tract, pancreas and sweat glands. Typically,
presentation is in infancy or childhood with recurrent pulmonary infections and
sometimes with malabsorption.
Cystinuria Screen
Specimen:
Urine mid-stream
Reference Range: Detected or not detected
Cystinuria with an incidence of 1:10,000 is one of the commonest of the inborn errors
of metabolism. Failure of the renal tubules to reabsorb cystine from urine results in
excretion of a high concentration of poorly soluble cystine which can precipitate to
form cystine stones 1–2% of all renal calculi.
Cytogenetics
Specimen:Amniotic Fluid
Bone marrow
Curettings from products of conception
Fetal tissue
Blood for karyotype – Whole blood, Lithium Heparin.
Collect specimens on a Monday – Thursday. Please inform the laboratory on
02 6285 9803 if collection is required out of these times.
Three major areas of testing are carried out:
1. Prenatal diagnosis from:
• Amniotic Fluid
• Chorionic Villi.
Capital Pathology Handbook – Interpretation of Laboratory Tests
2. Clinical diagnostic work for cases of:
• Infertility or Multiple Miscarriages
• Stillbirth (Where Clinical Indicators are Present)
• Babies With Multiple Malformations, With or Without Neurological Dysfunction
• Children With an Unusual Appearance Who Are Developmentally Delayed or
Mentally Retarded, Especially if There are Coexisting Congenital Defaults
• In Some Children With Speech Delay, Behavioural Problems, Isolated Mental
Retardation
• Genital Abnormality, Hypogonadism.
3. Cancer Cytogenetics:
• Leukaemia
• Lymphomas
• Solid Tumours.
Cytology
The Cytology Department at Capital Pathology is a fully accredited comprehensive
cytology laboratory, which processes a full range of gynaecological, non–gynaecological
and fine needle aspiration specimens. We participate successfully in the
RCPA External Quality Assurance programme and our Performance measures
are amongst the best in Australia.
See
Bronchial Specimens
Cerebrospinal Fluid (CSF)
Cervical Cytology (includes Pap smear and HPV testing)
Cyst Fluids
Effusion Fluids (Pleural, Ascitic, Synovial)
Fine Needle Aspirate (FNA)
HPV testing
Nipple Secretions
Sputum Cytology
Urine Cytology
C
A
Cytomegalovirus (CMV) Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Interpretation
IgM antibodies, reported as +ve or –ve, are detectable for a period of about 6 months
from the time of commencement of a CMV infection. False positives occur during
some other viral infections, notably those due to EBV.
IgG antibodies, reported in units/L, become detectable soon after the commencement
of infection and remain positive for life, usually at a level > 20 units/L. A rising titre in
paired sera is the best evidence of current infection. Most adults are IgG CMV positive
indicating previous infection.
The virus itself persists in latent form throughout life after recovery from the initial
infection and can be reactivated in an immunocompromised patient.
Clinical infection can present in several ways:
•
•
•
•
Subclinical infection is common, particularly in childhood.
A mononucleosis–like syndrome is found in adolescents and young adults, spread
by sexual and other intimate contact. CMV differs from EBV mononucleosis
in its absence of heterophile antibodies. Exudative pharyngitis and cervical
lymphadenopathy are rare. The illness can be severe with fevers and profound
fatigue lasting several weeks and the virus can cause hepatitis. As in EBV
infections, variant lymphocytes are a feature.
Congenital infections range from inapparent to severe with congenital
abnormalities or intrauterine death. Diagnosis is the identifying of the virus in urine
collected during the first week of life.
Immunocompromised patients can develop severe generalised disease.
Capital Pathology Handbook – Interpretation of Laboratory Tests
D
D
A
D–Dimer
Specimen:
Plasma – Sodium Citrate
Reference Range: < 0.2 mg/L
D–dimer is a fibrin break–down product and elevated levels can be found in any
situation where there is thrombosis.
Clinically the test is used when DVT (deep vein thrombosis) with or without PE
(pulmonary embolism) is suspected. A normal D–dimer level makes DVT/PE unlikely.
Elevated levels have a poor predictive value for DVT/PE as they are found in many
clinical situations including chronic inflammatory disorders, malignancy, post–operative
states and acute rheumatic conditions.
See
Coagulation Studies: Coagulation Screen
Dehydroepiandrosterone and Dehydroepiandrosterone Sulphate (DHEA, DHEAS)
Specimen:
Serum – Gel
Reference Range: Supplied with report
DHEA and its sulphate DHEAS are the most abundant androgenic steroids secreted
by the adrenal cortex. DHEAS is more commonly measured than DHEA and for
practical purposes the two estimations provide the same information.
Their uses are:
•
•
•
•
In the investigation of virilism – levels are usually > 3x upper limit of normal
In the investigation of hirsutism when there is a more than 2–fold increase in
free testosterone
To differentiate Cushing’s disease (minor elevations of DHEAS) from adrenal
neoplasms where there are large increases
For monitoring steroid suppression therapy in congenital adrenal hyperplasia.
Dementia (Acute or Chronic Organic Reaction)
The cardinal features are alteration in level of consciousness, confusion and focal
or global cognitive impairment. It is important to exclude treatable causes, and so
consideration can be given to the following lists as a guide.
Causes
• Cerebral lesion–infarct, haemorrhage, raised intracranial pressure, trauma,
abscess, tumour which could be primary or secondary
• Sepsis – consider septicaemia , bacterial endocarditis, subphrenic abscess,
meningitis, urinary tract infection, encephalitis
• Specific infections – HIV, neurosyphilis, Creutzfeldt-Jakob disease
• Metabolic – diabetes, acidosis, uraemia, decompensated liver disease,
hypothyroidism (myxoedma madness), Cushing’s Disease, hypercalcaemia
(psychic moans), hypoxia
• Drug reactions
• Hospitalisation in the elderly
• Acute or first presentation of psychosis / psychoaffective disorder
• Poisoning – mushrooms, prescribed medications in overdose, illicit drugs, lead,
ethanol
• Primary dementias and neurological degenerative disorders – Alzheimer’s
disease, multi-infarct demetia, Huntington’s chorea, storage diseases.
Tests to consider
• FBC and ESR
• CRP
• U&E, LFT, calcium, magnesium
• Vitamin B12 and folate
• Syphilis serology
• Thyroid function tests
• Lumbar puncture and CSF examination
• Blood cultures
• HIV serology
• Serum cortisol, or 24-hour urine cortisol
• Plasma glucose
• Blood gases
• MSU
• CXR, ECG, cerebral CT.
Dengue Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Dengue, a painful febrile illness (“break-bone fever”) caused by a mosquito-borne
arbovirus, is found all over the Pacific including Fiji. Typically it occurs in outbreaks but
can be sporadic.
Diagnosis is by measuring IgM and IgG antibodies preferably in paired sera. Although
both antibodies may date back to an earlier infection, the ratio of IgM to IgG, or rising
titres in paired sera, will help decide whether infection is recent.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Deoxypyridioline Cross Links (DPD)
Specimen:Urine – Collect the second morning void into
urine container (nil preservative).
Reference Range:Supplied with report
Useful bone turnover marker.
Desipramine (Pertofiren)
Specimen:
Plasma – Lithium heparin
Reference Range: Supplied with report
See
Antidepressant Drugs, tricyclic
Dexamethasone Suppression Test
By arrangement with collecting rooms or main laboratory
Specimen:Serum – Gel
Protocol:
Day 1, collect baseline cortisol specimen at 08.00-09.00 hours.
Give 1 mg dexamethasone orally at 23.00 hours
Day 2, collect cortisol specimen at 08.00–09.00 hours
Reference Range: Cortisol < 100 nmol/L at 09.00 hours post–suppression
Normal suppression, especially to < 50 nmol/L makes Cushing’s syndrome unlikely.
Failure to suppress is less helpful and may occur with a range of conditions including:
•
•
•
•
•
Cushing’s syndrome
Endogenous depression
Chronic alcoholism
Drugs–phenytoin, barbiturates, oestrogens
Significant stress or illness.
D
A
Diabetes insipidus
The essential feature is inadequate ADH (antidiuretic hormone) from the
hypothalamus/posterior pituitary causing excessive loss of dilute urine from the
kidney; or a kidney which is unresponsive to ADH. The usual problem is to separate
compulsive water or simple urinary frequency from true diabetes insipidus in a patient
complaining of polydipsia and polyuria.
•
•
•
•
•
ther causes of polyuria need to be eliminated.
O
diabetes mellitus
hypercalcaemia
renal disease
medication: lithium, diuretics.
A 24–hour urine should be collected to check whether urine volume is genuinely
excessive e.g. > 4 L. in severe disease, volume can be as high as 18 L.
The final screen test is urine osmolality after 8–12 hours of water deprivation
overnight. A urine specimen is collected on waking and 1/2–1 hour later before
drinking anything, the osmolality in these two specimens indicating ability to
concentrate urine.
The person with diabetes insipidus will excrete a large volume of dilute urine. Water
restriction should only be performed under medical supervision, due to the risk of
serious dehydration.
Diazepam
Specimen:Plasma – Lithium heparin tube
Trough level is taken before next dose (within 1 hour).
Reference Range: Supplied with report
Dibucaine Number
Specimen:
Serum – Gel
Reference Range: Supplied with report
The dibucaine number detects the qualitative difference in cholinesterase enzymes in
scoline–sensitive persons.
SeeCholinesterase
DIC (Disseminated Intravascular Coagulation)
This is an uncommon condition in which there is a generalised consumption of plasma
clotting factors and platelets resulting in fibrin deposition within the microcirculation.
Secondary haemorrhagic events are due to the consumption of normal clotting factors
and secondary fibrinolysis.
Clinical setting for DIC:
•
•
Major trauma
Septicaemia (usually with acidosis)
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
•
Obstetric crises (placental abruption, eclampsia, retained dead fetus)
Malignancy (acute promyelocytic leukaemia).
Tests for DIC include FBC (with blood film for fragments), platelet count (as part of
FBC), prothrombin time, APTT, fibrinogen assay and D–dimers.
Digoxin
Specimen:Serum – Gel
Specimen should be collected at least 6 hours post dose.
Therapeutic Range:0.6–1.3 nmol/L
Dilantin
Specimen:Serum – Gel
Tough level is taken just before next dose ( less than one hour).
Peak level is collected between 4–7 hours post dose.
Reference Range: 40–80 umol/L therapeutic range adults
Disaccharidases
SeeHistopathology
Disopyramide (Rythmodan)
Specimen:
Plasma – Lithium heparin
Reference Range: Supplied with report
DNA Antibodies (Anti–double Stranded DNA)
Specimen:
Serum – Gel
Reference Range: Supplied with report, qualitative test
Interpretation:< 1:20 or 1:40 equivocal
> 1:80 supports diagnosis of SLE
Anti–ds DNA is positive at 1:80 or higher in 60–80% of SLE. Low titres may be seen in
rheumatoid arthritis, autoimmune hepatitis and in other immunological disorders.
Dothiepin (Prothiaden)
Specimen:Plasma – Lithium Heparin
Suggest take a trough level just before next dose (within one hour).
Reference Range: Supplied with report
See
Antidepressant Drugs, tricyclic
D
A
Down’s Syndrome Screening
See
Prenatal Testing
Doxepin (Sinequan)
Specimen:Plasma – Lithium Heparin
Suggest take a trough level just before next dose (within one hour).
Reference Range: Supplied with report
See
Antidepressant drugs, tricyclic
Drug Screen, Screen for Drugs of Abuse
Specimen:
Urine – Random
Reference Range: Detected or Not Detected
Tests for drugs of abuse
Qualitative drug screening may be undertaken to determine whether a person has
taken a medication or drug. Testing is useful to ascertain:
•
•
•
Compliance with prescribed drugs e.g. methadone
Use of non–prescribed drugs
Accidental or deliberate abuse of illegal drugs.
Factors affecting a valid specimen
Urine creatinine concentration is assayed to detect diluted specimens. This can
be caused by surreptitious addition of tap water or excessive fluid intake prior to
collection. A urine creatinine level < 2.0mmol/L indicates a dilute specimen. A repeat
specimen should be considered.
The temperature of the specimen indicates if it is freshly passed. If the specimen is
fresh, the temperature should fall in the range of 33–38’C. If the temperature is not in
this range, specimen substitution should be suspected.
Adulteration can affect the final results, and pH, smell and visual checks are all
performed to check for these possibilities.
Factors affecting detection time:
• Usage pattern
• Drug and dose of drug used
• Urine concentration
• Assay method and cut off value used.
Chain-of-custody
Urine samples are collected under supervision into specially designed beakers
with temperature strips affixed. The supervising collector is specially trained in the
requirements surrounding urinary drug screening and he/she records the temperature
of the specimen and divides the specimen into three tubes. Each tube is labelled and
signed by the client to verify identity. The tubes are then individually packaged with
tamper evident tape and placed in secure tamper-proof satchels for transport to the
laboratory.
Capital Pathology Handbook – Interpretation of Laboratory Tests
An audit trail is maintained such that every person involved in the collection, transport
and checking process is required to sign either the chain-of-custody form or the
transport form accompanying the specimens. The integrity of the samples and
transport satchel is noted on the forms throughout the process. These forms are
stored in a secure facility for future reference if required
Interpretation of Results
The presence of each drug or metabolite is tested for at or above a predefined cut-off
level. These levels are dictated by International Standards for urine drug testing and
defined in the Australian/New Zealand Standard AS/NZS 4308:2008. The “cut-off” levels
are established because the aim of workplace testing is usually to identify significant
residues of the targeted drug, not minute traces. For a result to be “non-negative”, the
amount of the drug detected must be at or above the “cut-off” level. If a drug is detected
but the level is below the “cut-off” the result will be negative. The “cut-off” levels for some
classes differ for screening and confirmation. This is due to the non-specific nature of
the screening assay versus the highly specific nature of the GC/MS confirmation.
A confirmed positive result reveals the presence of a drug in the specimen at or above
the “cut-off” level. It gives no information about how or when the drug was taken.
It also does not provide an indication of impairment. A positive result may relate to
previous drug use with no current physical effects. Positive results are reported as
‘Detected’, while a negative result is reported as ‘Not Detected’.
Drugs Detectable
The general screen detects groups of drugs including: opiates; methadone;
amphetamines; cocaine; benzodiazepines; and cannabis. Further confirmatory testing
can be undertaken on request.
Please note Capital Pathology collection centres have accredited staff who have
completed the ASNZ4308:2008 training.
D–Sialated Transferrin Assay
Specimen:
Collect nasal / fluid drips into sterile container
Reference Range: Supplied with report
Used for identification of CSF in a discharge fluid.
D
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
E
Ear Swabs and Infections
Otitis media
It is not possible to sample the middle ear routinely and the flora of the external
meatus bears no relation to that behind the drum. Antibacterial treatment is therefore
empirical, based on studies which show the role of H. influenzae, Strep. pneumoniae,
Moraxella (previously Branhamella) catarrhalis, and Strep. pyogenes. Amoxycillin,
cotrimoxazole and amoxycillin–clavulanate provide effective treatment. Cefaclor
has insufficient activity against Strep. pneumoniae for it to be recommended in this
situation.
Otitis externa
Ear swabs are taken from just inside the external meatus. The commonest pathogens
are Staph. aureus and pseudomonas aeruginosa. H. influenzae and S. pneumoniae
are also frequently isolated.
ECGs
These are performed at Capital Pathology Collection Centres. With a request for
“ECG Trace only” you will be provided with the ECG for your own interpretation. With
a request for “ECG Trace and report” you will be provided with the ECG and a report
from a cardiologist.
Please indicate on the request form when an ECG is urgent.
Holter Monitor services are also provided by Capital Pathology, with a report and
interpretation from a cardiologist.
Please contact your most convenient Collection Centre to arrange an appointment.
Effusion Fluids (Pleural, Ascitic, Synovial)
Effusion fluids should be placed into a labelled specimen container and refrigerated at
4ºC. Storage overnight is satisfactory. It is preferable to send the whole specimen with
a minimum of 100 mL, to allow for concentration of poorly cellular specimens and for
preparation of a cell block. The cell block is then available for special staining including
immunohistochemistry should this be necessary for the diagnosis.
E
A
eGFR (Glomerular Filtration Rate)
GFR can be calculated in a number of ways. The MDRD formula has now become
available to calculate estimated GFR (eGFR) based on serum creatinine, age and sex.
eGFR is routinely reported on patients aged 18 years and over. The aim of reporting
eGFR is to detect kidney disease at a time when there may be benefit from therapy
to prevent complications. There are a number of limitations that apply to using the
formula. The formula has not been fully validated in certain populations (Aboriginal,
Asian and Torres Strait Islander), and is not appropriate for pregnancy, obesity,
dialysis, rapidly changing renal function or with some diets (vegetarian).
Full details of the application of eGFR can be found in the position statement from
Medical Journal Of Australia 2005 Vol 183:3 pp138–142.
Electrophoresis of Serum Proteins
Specimen:
Serum – Gel x 2
Reference Range: Supplied with report
Serum protein electrophoresis (EPP) is an essential test when interpreting a raised
total globulin or immunoglobulin to determine whether a paraprotein is responsible for
the increase.
A. Monoclonal bands, paraproteins, M–bands
A paraprotein, which appears as a sharply defined abnormal band on serum EPP,
consists of a single population of identical immunoglobulin molecules formed by a
clone of neoplastic plasma cells. They can be benign (MGUS) or malignant (myeloma,
macroglobulinaemia or lymphoma).
Occasionally an M–band is due to an extra–medullary plasmacytoma.
Benign paraproteins (MGUS – Monoclonal Gammopathy of Uncertain
Significance)
These paraproteins, composed of IgG, IgA, or IgM, appear with increasing frequency
in the elderly – 8% of patients over age 70. They can be transient, particularly
in younger individuals. Characteristics are a relatively low concentration (total
IgG < 20g/L, IgA or IgM < 10g/L), normal blood picture, no depression of normal
immunoglobulins, no urine Bence Jones Protein. Bone marrow examination is not
usually indicated but if done will show < 10% plasma cells with no atypical cells.
MGUS can be regarded as a “carcinoma in situ” of the immune system and like any
in situ lesion it has the capacity to progress to malignancy, quickly or slowly, or it can
remain largely static for the life of the individual, hence the need for monitoring at least
annually. Evolution to multiple myeloma (or macroglobulinaemia) occurs in 25% of
patients.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Malignant paraproteins
These consist either of IgG or IgA (multiple myeloma) or IgM (Waldenstrom’s
macroglobulinaemia or lymphoma) and show the invasive characteristics of
malignancy with infiltration and destruction of bone marrow.
Characteristics of a malignant, as compared with a benign, paraprotein are:
•
•
•
•
•
•
•
•
Anaemia, thrombocytopenia, neutropenia
Immune paresis, i.e. reduction of normal IgG, IgA and IgM
High and rising concentration of paraprotein
Bence Jones protein (free light chains) typically appears in the urine in myeloma
Myeloma causes diffuse osteoporosis or lytic lesions, either of which can cause
back pain, compression fractures or bone pain in other parts of the body such as
ribs or pelvis
Hypercalcaemia, raised ALP
Renal impairment
Hyperviscosity syndrome, particularly with macroglobulinaemia.
Bone marrow examination and skeletal survey establish the diagnosis.
Where there is a strong clinical suspicion of myeloma (pancytopenia, osteolytic
lesions, bone pain, immune paresis) but no serum M–band, urine EPP may show
Bence Jones myeloma in which the band (of free light chains) is found only in urine.
Polyclonal increases
These are reflected in serum proteins as an increase in total globulins and in the EPP
as an increased density in the gamma zone.
They are caused by any chronic inflammatory or liver disorder:
•
•
•
•
Chronic hepatitis, viral, alcoholic or autoimmune
Connective tissue disease
Chronic inflammatory bowel or pulmonary disease
Parasitic infestations.
Electrophoresis of urine
Specimen:
Random urine
Endomysial Antibodies (EMA)
Specimen:
Serum – Gel
Reference Range: Supplied with report
EMA is no longer recommended for coeliac disease diagnosis, as TTG is the preferred test.
See
Coeliac Disease
E
A
Eosinophils
Specimen:
Whole blood – EDTA
Reference Range: Adults < 0.6 x 109/L
Eosinophilia
Eosinophil counts are part of a routine blood count. The common causes of
eosinophilia are drug effect, allergy or parasitic infestation of the gut.
The differential diagnosis includes:
•
•
•
•
•
•
•
•
•
Allergic disorders
Parasitic colonisation – hookworm, filaria, hydatids, toxocara
Drug administration
Skin disease, e.g. eczema, psoriasis, pemphigus
Collagen disorders, especially polyarteritis nodosa
Infections, e.g. TB, scarlet fever
Malignant disease, e.g. lymphoma, Hodgkins disease, ovarian cancer
Pulmonary eosinophilia
Hypereosinophilic syndrome (a rare myeloproliferative disorder).
Epilim (Valproate)
Specimen:Serum – Gel
Suggest trough level collected just before next dose. If peak
requested collect between 0.5–1.0 hours for Syrup, 1–3 hours
for capsules and 2–6 hours for coated tablets.
Reference Range: Therapeutic 350–700 umol/L
Epstein-Barr Virus Antibodies
Specimen:
Serum for heterophile antibodies, liver enzymes, EBV antibodies
Whole blood – EDTA for blood count.
Epstein–Barr Virus is the causative agent in infectious mononucleosis which is
characterised by lymphadenopathy, pharyngitis, fever, variant lymphocytosis in the blood
film and transient heterophile antibodies in serum. Fatigue can be profound and continue
for months.
EBV antibodies
Where EBV infection is suspected but heterophile antibodies remain negative–as
happens in 10% of young adults, and 50% of children – the measurement of EBV
antibodies will provide a definitive answer.
VCA (Viral Capsid Antigen) IgM antibodies appear at 4–7 days after symptoms
develop and persist for 2–4 months, occasionally up to 1 year. Their presence
indicates current or recent infection.
VCA IgG antibodies appear at the same time as IgM but persist for life.
EBNA (Anti EB Nuclear Antigen) antibodies appear 3–6 weeks after onset and
also persist for life. 80% of the population are EBNA +ve, indicating past infection.
A minority of people infected with EBV never develop EBNA antibodies but will have
a positive VCA IgG test to indicate their past exposure.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Erythropoietin (EPO)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Erythropoietin (EPO), a hormone produced by the kidney in response to low renal p02,
has an important role in the regulation of erythropoiesis.
Measurement of EPO can be used when differentiating primary polycythaemia (low
EPO) from secondary polycythaemia (high EPO).
Reduced EPO formation is a feature of the anaemia of chronic renal disease and EPO
can be used to treat the anaemia.
Erythrocyte Sedimentation Rate (ESR)
Specimen:
Whole blood – EDTA
Reference Range:
Age
Child 1–18
female
< 16
mm/hr
male
< 16
Adult < 50yrs
< 21
< 16
Adult > 50yrs
< 36
< 31
Pregnancy
< 100
Levels are higher in pregnancy due to hyperfibrinogenaemia.
The ESR is an antique test that still has a place as a nonspecific indicator of
inflammatory disease and abnormal protein states.
In an acute illness, the ESR may take a week or more to start to rise and stay elevated
for some weeks after its resolution. The CRP, by contrast, rises and falls more quickly
and is in some ways a better marker for acute inflammation.
See
C – Reactive Protein (CRP)
Essential Thrombocythaemia
Essential thrombocythaemia is a myeloproliferative disorder characterised by a
sustained increase in the platelet count, particularly above 600 x 109/L.
Clinical features include splenomegaly, an increase in thrombotic risk and bleeding of
varying severity. Bone marrow biopsy and elevated NAP score are useful in diagnosis.
Treatment is aimed at reducing the platelet count. Therapy at counts which are only
modestly increased is controversial.
Ethosuximide (Zarontin)
Specimen: Plasma – Lithium heparin
Trough level is taken before next oral dose ( within one hour).
Peak level 2–4 hours post dose.
Reference Range: Supplied with report
E
A
Extractable Nuclear Antigens (ENA)
Specimen:
Serum – Gel
Reference Range: Supplied with report, qualitative test
Some nuclear antigens can be extracted into solution and patients’ serum tested for
presence of antibodies against those antigens. ENA screen is used as a follow–up
test on positive ANAs or where one of the connective tissue diseases other than
rheumatoid arthritis is suspected.
Antigen
Associated disease
(with approximate % frequency of antigen)
Sm
SLE (20%)–antigen is 99% specific when present
RNP
MCTD
Autoimmune hepatitis
SLE (30%)
RA
Ro (SS–A)
Sjogren’s (90%) usually associated with La (SS–B)
SLE (40%)
Polymyositis (5%)
RA (5%)
Neonatal lupus in pregnancy
La (SS–B)
Sjogren’s (80%) usually associated with Ro (SS–A)
SLE (10%)
Diffuse scleroderma
Neonatal lupus in pregnancy
Scl 70
Diffuse scleroderma (50%)
CREST (10%)
Jo1
Polymyositis (30%)– > 95% specific when present
Eye Swabs
If the eye is moist, use a dry swab; if the eye is dry, moisten the swab in transport
medium. After pulling the lower lid down roll the swab across the inner part of the
lower lid. If there is pus in the corner of the eye, get some of this onto the swab as
well.
In neonates, conjunctivitis can be due to gonococci or Chlamydia trachomatis
transmitted from mother to baby during birth. When testing for Chlamydia remove any
purulent exudate before collecting conjunctival epithelial cells by rubbing the small
Chlamydia swab over the everted palpebral conjunctiva.
Bacterial conjunctivitis outside the neonatal period is most commonly caused by Staph.
aureus, H. influenzae or Strep. pneumoniae and usually resolves spontaneously or in
response to topical eye drops or ointment. Occasionally unusual bacteria are found.
A negative bacterial culture usually indicates a viral or allergic conjunctivitis.
Capital Pathology Handbook – Interpretation of Laboratory Tests
F
F
A
Factor Assays (Coagulation Factor Assays)
Specimen:Plasma – Sodium Citrate (2 x 4.5 mL)
Sodium Citrate tubes must be filled to capacity.
Spin, separate and freeze plasma.
Reference Range: Supplied with report
See Coagulation Studies: Factor Assays
Factor V Leiden (R506Q)
Specimen:
Whole blood – EDTA
The most common of the inherited causes of hypercoagulability, first recognised in
1993, is a mutation of the Factor V gene known as Factor V Leiden. Normal Factor
V, which promotes normal clotting, is inhibited physiologically by Activated Protein C.
Factor V Leiden is abnormally resistant to this inhibitory action and the presence of
Factor V Leiden is detected by the APCr test.
Factor V Leiden occurs in 3– 7% of Caucasians and is a factor in 20–40% of cases of
venous thromboembolism (VTE). Risk of VTE is increased 5–10 times in heterozygotes
compared with normal people and 5–100 times in homozygotes. Addition of oral
contraceptives increased the risk a further 3–5 times in heterozygous individuals.
Faeces, for Clostridium Difficile Toxin
Specimen:Faeces
Note any antibiotic therapy.
Reference Range: Not detected
Faeces, for culture and microscopy
Faeces Examination for Microbiology
Specimens should be fresh and submitted contained in the brown top faeces jar.
Optimum sensitivity in stool examination is provided by the permanent fixed smear
which is routinely performed at Capital Pathology. The standard faeces series is,
according to best clinical practice and current HIC tables, two faeces specimens
within a seven day period. Microscopy and culture is performed on specimen one
and microscopy alone is performed on specimen two.
If a specimen of faeces is unobtainable, a rectal swab may be used (in transport
medium). This not adequate for Clostridium difficile toxin detection or for parasites.
Please specify if non–routine examinations are required, (e.g. Rotavirus, Clostridium
difficile, Viral culture).
Patient Information leaflets are available from the Client Services Department on
02 6285 9802 or at any Collection Centre.
Faeces, for Occult Blood
Specimen:Special diet required – please contact Collection Centre for
information leaflets. Collect samples on three separate days.
Indications
When done properly, a positive result (even one) increases likelihood of
gastrointestinal blood loss, especially from lower bowel.
However, because of low sensitivity for detecting small, often intermittent bleeding,
a negative result does not exclude GI blood loss as from a tumour. Further evaluation
(e.g. colonoscopy) is still therefore indicated in at-risk patients with unexplained iron
deficiency.
Types of test/dietary advice
Nonspecific guaiac tests
These, the most commonly used tests, detect the peroxidase activity of haem whether
of human or animal origin.
They can give false positives with high meat diets, peroxidase–containing foods such
as horseradish or turnip, or sometimes with iron supplements. Gastric irritant such as
NSAIDs can give positives because of the minor bleeding they induce.
High dose vitamin C or E supplements can give false negatives.
Specific immunochemical tests
Most of these detect intact human Hb but others are based on human haem, porphyrin
or even albumin. Although these tests avoid dietary false positives, there is still the
large problem of positives due to minor benign bleeding.
Faeces, for pH and Reducing Substances
Specimen:Faeces
Require same day specimen.
Reference Range: Supplied with report
Faeces, for Rotavirus
Specimen:Faeces
Reference Range: Not detected
Ferritin
Specimen:
Serum – Gel
Reference Range: Adult female < 50 years
Adult female > 50 years
Adult males
15–200 ug/L
30–300 ug/L
30–300 ug/L
Capital Pathology Handbook – Interpretation of Laboratory Tests
Fibrinogen
Specimen:
Plasma – Sodium Citrate
Reference Range: 2.0–4.0 g/L
Fibrinogen is reduced in:
•
•
•
Afibrinogenaemia (total absence of fibrinogen)
Hypofibrinogenaemia (decreased synthesis of fibrinogen)
Dysfibrinogenaemia (structural abnormality of the fibrinogen molecule).
Fibrinogen is increased in inflammatory conditions.
It is measured as part of the basic haemostasis screen.
Filariaria Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Filariasis occurs in Africa, Latin America, Asia and the Pacific Islands where the
most common cause is Wuchereria bancrofti. Diagnosis is by finding microfilaria
in peripheral blood. An eosinophilia may be the only evidence of filarial infestation.
Lymphatic filariasis occurs when host inflammation and fibrosis lead to lymphatic
occlusion and in patients heavily and repeatedly infected, elephantiasis may develop.
Bacterial cellulitis contributes to tissue damage. The syndrome of tropical eosinophilia
is an extreme reaction to filariasis.
Treatment with diethyl carbamazine or ivermectin clears microfilaria from the blood.
Dying microfilaria can stimulate a severe allergic reaction requiring treatment.
Fine Needle Aspirate (FNA)
FNA is an easy, relatively painless and inexpensive procedure, which should be
considered in the diagnostic work–up of any palpable, non–pulsatile mass. The
complication rate is low, with local haematoma formation and bruising the most
commonly occurring complications.
FNA has high sensitivity (i.e. low false negative rate) and high specificity (i.e. low false
positive rate). The specificity of FNA approaches that of frozen section analysis and in
experienced hands, has a false positive rate of less than 0.5%.
The most common site for FNA is the breast, but thyroid nodules, head and neck
lumps, enlarged lymph nodes and soft tissue masses can all be aspirated.
The procedure will be performed by one of Capital Pathology’s experienced
cytopathologists. Alternatively, specimens obtained by the referring doctor can be sent
to the laboratory via the courier for processing.
F
A
Appointments
Please phone the Cytology Department to arrange a patient appointment on
02 6285 9867. A patient information sheet is available from the Client Services
Department on 02 6285 9802, and this also has a map to direct the patient to the
laboratory.
Procedure
FNA is performed without anaesthesia using a 23 or 25 gauge needle. Negative
pressure is sometimes applied by retracting the plunger of an attached syringe while
moving the needle backwards and forwards within the lesion.
Cytological assessment is optimised by preparing both wet-fixed and air-dried
smears. Wet-fixed slides should be fixed as rapidly as possible using spray fixative
or by immersion in 95% alcohol for 20 minutes. The slides then can be air-dried and
placed in a slide mailer. When making air dried smears, complete and rapid drying
is necessary before transportation. A hair dryer on low setting can be used. Slide
smearing techniques can be discussed with the pathologists. It is necessary to write
on the frosted end of the slide whether that particular slide has been wet-fixed (WF)
or air-dried (AD), as the laboratory staining for each of these techniques is different.
The needle should be washed out using RPMI. The washings are submitted for
preparation of a cell block. Two or more aspirations will generally result in a greater
yield of cellular material.
A separate pass transferred into RPMI is advisable when investigating enlarged
lymph nodes or unusual masses. This allows for lymphocyte marker studies by flow
cytometry or for immunohistochemical staining,used in the investigation or tumours
of uncertain lineage.
Smear preparation
For non–gynaecological and fine needle aspiration specimens (see diagrams)
The simplest way to make a smear is the “pull apart method”. This is particularly useful
for bloody specimens and cyst fluids:
A small drop of fluid is placed on a clean glass near the label end. A second slide is
placed parallel over the first with the label end of the second slide at the opposite end.
Let the fluid start to spread between the two slides and then gently pull them apart
The specimen usually distributes relatively evenly between the two slides.
The second method is useful for thick fluids or aspirates. Much of the specimen
remains on the test slide, leaving the spreading slide relatively clean. All slides should
be sent to the laboratory.
Fix rapidly, by spraying with pump fixative or dropping into 95% alcohol, or by rapidly
air drying. Both wet–fixed and air–dried slides are useful, so it is preferable to use
both methods if the amount of material is sufficient. However it is important to label the
slides as to which method has been used.
Please ensure that the slides are labelled with pencil on the frosted end, with the
patient’s full name and date of birth.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Slide preparation diagrams
Right angle
for thick material
Pull apart
for bloody or cystic material
To discuss any matter related to FNA with a pathologist, please phone 02 6285 9867.
Results are usually available the following day.
An FNA Collection Procedure Sheet is available from the Client Services on
02 6285 9802 on request.
F
A
Yellow top jar
Available from
laboratory.
For suspected
lymphomas
RMPI-FCS
Spray fixed
(Pap stain for nuclei)
Cell blocks helpful especially
if primary tumour unknown
or to prove a metastasis
from a known tumour.
Immunohistochemistry for
tumour characterisation
possible.
Flow cytometry helpful in
classifying NHL
A separate pass into the
RPMI is recommended as it
gives more material
Alcohol spray fixative.
Hair dryer helpful.
Made at the time of
FNA – BOTH methods of
preparation preferred for a
single episode.
Direct Smears
Air dried
(Romanowsky stain)
Comment
Specimen Type
Cell blocks – if primary
tumour unknown or
to prove metastasis
from unknown tumour.
Immunohistochemistry
for tumour
characterisation
possible.
Cytology of solid
lesion
Flow cytometry
helpful in
classifying NHL
For suspected
lymphoma
Test Required
Cytology
of fluids
Contact
laboratory if
special tests
required
Microbiology/
culture
Fine Needle Aspiration Specimen Handling Flow Chart – 2012
Capital Pathology Handbook – Interpretation of Laboratory Tests
F
A
Flow Cytometry
Specimen:
Whole blood – EDTA x 2
This may also be performed on fresh, unfixed lymphoid or other tissue removed
surgically or by fine needle aspiration. Blood or bone marrow aspirate may be
submitted to flow cytometry.
This test is used for the classification of lymphomas and leukaemias and to establish
a diagnosis of reactive or neoplastic lymphoid proliferations.
The report will include quantitation of cell lines identified.
See
Lymphocyte Markers
Flucytosine (Ancotil)
Specimen:
Plasma – Lithium Heparin
Sample just before next dose and 2 hours after oral dose
(or 30 minutes after IV dose).
Reference Range: Supplied with report
Fluid Examination
Body fluid
Specimens should be transported in a yellow top jar or as a swab in transport medium.
Depending on the clinical circumstances individual requests should specify which tests
are required e.g. microbiology, cytology or biochemistry (protein and glucose).
Cerebrospinal fluid
See Cerebrospinal Fluid (CSF)
Peritoneal dialysis
Send entire bag of peritoneal dialysate. Usually one bag per 24 hours is required.
Pus
Where sufficient pus is available, aspirate with a sterile syringe fitted to a broad gauge
needle. Inoculate some of the pus into a blood culture bottle and leave the remainder
in the syringe. Remove air from the syringe and carefully remove the needle and
discard appropriately. Cap syringe and send to the laboratory.
Synovial fluid
For full synovial fluid analysis the specimen should be apportioned as follows:
1 EDTA for cell count. Collect 2 mL, mix immediately and thoroughly.
1 sterile specimen container for crystals, gram stain, culture, and urate level.
Folate, red cell
Specimen:
Whole blood – EDTA
Reference Range: > 1000 nmol/L
Red cell folate is a better indicator of tissue stores than serum folate, which is more
labile.
Interrelationship between B12 and folate deficiencies:
1° deficiency
Serum B12
Serum folate
Red cell folate
B12

n
n
Folate
n


Follicle Stimulating Hormone (FSH)
Specimen:
Serum – Gel
Reference Ranges:
Adult female
FSH IU/L
Follicular and luteal phases
2– 23
Mid–cycle ovulatory peak
11– 30
Post–menopausal
21– 175
Adult male
FSH IU/L
1.0– 14
Applications
Identifying the menopause – the FSH rises, FSH rises relatively more than LH.
Ovarian or testicular failure – both FSH and LH rise to menopausal levels. Gonadal
failure can be primary as in agenesis, dysgenesis, Klinefelter’s or Turner’s syndromes.
Or it can be secondary as in premature menopause, oophorectomy or testicular
damage.
Identifying day of ovulation–in fertility control.
Hypopituitarism – LH and FSH are not usually undetectable in hypopituitarism but
low or inappropriately normal values in patients with low oestradiol (women) or
low testosterone (men) are suggestive. Mid-range or high values can help exclude
hypopituitarism.
Polycystic Ovary Syndrome–LH is often raised relative to FSH with an
LH/FSH ratio > 2.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Fructosamine
Specimen:
Serum – Gel
Fructosamine is useful in the monitoring of diabetes mellitus and the assessment of
overall control of blood glucose over the one to three weeks prior to collecting the
specimen. Increased values are suggestive of significant persistent hyperglycaemia.
Levels are lower in the presence of hypoalbuminaemia. It is useful together with the
assay of haemoglobin A1C (glycated haemoglobin) which reflects diabetic control over
a longer period.
SeeHbA1c
FTA (Fluorescent Treponemal Antibody)
Specimen:
Serum – Gel
Reference Range: Not detected
A positive FTA indicates past or present treponemal infection. It does not distinguish
between syphilis and yaws. The FTA stays positive life–long.
See Syphilis
Fungal Investigation
Skin lesions – take a scraping of the advancing edge.
Nails – nails should be cut back. Collect clippings or scrapings, including necrotic
debris from beneath the nail.
Hair – include hair roots.
Collect specimen into a yellow top jar. The more material submitted the greater the
odds of positive findings.
Microscopy is reported within 24 hours. Cultures are maintained and examined on a
regular basis for 2–4 weeks.
F
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
G
G
A
Galactorrhoea
Most patients are women but occasionally galactorrhoea is found in men.
Causes are:
•
•
•
Hyperprolactinaemia
This is commonly, but not invariably, associated with galactorrhoea. There may
or may not be a pituitary tumour. Elevations of prolactin can be minimal and
intermittent.
Hypothyroidism and hyperthyroidism
Drugs
phenothiazines, benzodiazepines, tricyclics, methyldopa, butyrophenones,
metoclopramide, H2 receptor antagonists, digoxin, spironolactone, post–oral
contraceptive.
Galactose
Specimen:Random urine
Specimen needs to be frozen.
Reference Range: Supplied with report
Gallstones
These consist mainly of cholesterol except in chronic haemolysis when pigment
(bilirubin) stones may be formed. Analysis of a stone is of interest only when
haemolysis is suspected or when an unidentified object ?gallstone, is found in faeces.
Gamma Glutamyl Transpeptidase (GGT)
Specimen:
Serum – Gel
Reference Range:Adult female
Adult male
5–35 U/L
5–50 U/L
GGT is found almost entirely in the liver. It is elevated particularly in cholestatic
disorders, by alcohol, and also as an effect of some drugs, notably anticonvulsants.
GGT is the enzyme most reliably raised by excessive alcohol, typically in the range
60–200 but occasionally > 1000. 30% of heavy drinkers have normal levels. After a
weekend binge, levels rise up to 100% over 3 days. If the cause of an elevated GGT is
uncertain, total abstinence from alcohol with weekly GGT estimations over a 4–week
period may provide the answer.
See Liver Function Test / Interpretation
Gardnerella Vaginalis
Formerly known as Haemophilus vaginalis and Corynebacterium vaginalis.
A common bacterial isolate from the vagina. It may be present in up to 30% of
healthy women. When found in association with “non–specific vaginosis”, treatment
with metronidazole or tinidazole will eliminate odour and discharge though there is a
tendency to recurrence requiring repeat treatment.
See Vaginal Swabs for Discharge
Gastrin
Specimen:Serum – Gel
Patient should be fasting.
Spin, separate and freeze immediately.
Reference Range: Supplied with report
Gastrin, released from the gastric mucosa, is a potent stimulant of gastric acid secretion.
Gastrin measurements are used in the diagnosis of pancreatic gastrinomas associated
with severe peptic ulceration in the Zollinger–Ellison syndrome in which there is
marked hypersecretion of gastric acid.
Vagotomy, non–fasting state, renal failure, or the gastric acid hyposecretion due to
pernicious anaemia and chronic atrophic gastritis, are associated with an increase in
gastrin levels.
Gentamicin
Specimen: Serum – Gel
If Gentamicin is given as a single daily dose a measurement of plasma concentration
should be made between 6–14 hours after the end of the infusion. For intermittent
dosing regimes, peak or trough levels may be measured.
Gentamicin dose adjustment is required to avoid the effects of gentamicin toxicity.
Monitoring of renal function – renal function should be checked at regular intervals
during gentamicin treatment.
Specialist advice may be required for monitoring and assessment of gentamicin levels.
Giardia
Specimen:
1–2 random faeces samples
Giardia lamblia is a protozoan infesting the upper small bowel where it can cause
acute or chronic diarrhoea and sometimes fat and vitamin malabsorption.
It is commonly taught that three stool specimens are required to detect intestinal
parasites such as Giardia but several studies have shown that > 85% of cases are
detected on the first specimen. When evaluating chronic or relapsing diarrhoea it may
be necessary to send multiple specimens for testing.
Capital Pathology Handbook – Interpretation of Laboratory Tests
The two commonly–used tests for Giardia:
•
•
Detection of Giardia antigen in faeces using an Elisa method.
Microscopy for cysts in faecal concentrate. Occasionally trophozoites are seen in
acute infections.
The antigen method is the more sensitive.
Treatment with tinidazole or metronidazole is usually effective but may need to be
repeated.
Gilbert’s Syndrome
This common, harmless, inherited condition, in which the liver has reduced ability to
conjugate bilirubin, was first described in 1901 by the Frenchman Gilbert whose name
is usually, though not always, pronounced in the English fashion.
It affects somewhere between 2 and 10% of the male population depending whether
the cut–off is taken as 25 or 20 umol/L. The male/female ratio is about 4:1.
Because a genetic test is not available, diagnosis is based on exclusion.
Features are:
•
•
•
•
•
•
Elevation
of bilirubin is the only laboratory abnormality and the level seldom
exceeds 80 ug/L
Liver enzymes are normal
The bilirubin is unconjugated
There is no evidence of haemolysis as measured by reticulocytes and
haptoglobins
The bilirubin level fluctuates; the condition is often first detected in the 2nd–3rd
decade
Fasting, dehydration, fever, sea–sickness, intercurrent illnesses, can all cause the
bilirubin to rise, sometimes to the point where it becomes visible as jaundice – at
this point the patient may note malaise, blaming it on the Gilbert’s rather than the
precipitating factors.
Gliadin (gluten) Antibodies (AGA)
Specimen:
Serum – Gel
Reference Range: Supplied with report
See
Coeliac Disease
Glomerular Basement Membrane Antibodies
Specimen:
Serum – Gel
Reference Range: Not detected
G
A
Glucose
Specimen:Plasma (Fluoride Oxalate) is ideal, or serum if cells can be
separated within 2 hours
Reference Range:
Age
mmol/L
Fasting glucose
All
3.4– 6.0
Random glucose
Day 1
1.7– 5.6
Day 1– 2 yr
2.5– 5.6
> 2yr
3.4– 7.7
If the specimen is collected in a plain tube the glucose level will tend to be low by an
unpredictable amount because red cells consume glucose, causing the serum level to
fall. Fluoride prevents this glycolysis.
Diagnostic Criteria For Diabetes
Single fasting and symptoms
2 hours post prandial or
casual post prandial and symptoms
> 7.0 mmol/L or
> 11.1 mmol/L
If no symptoms or equivocal symptoms:
At least one additional glucose measurement (preferably fasting) on a different day
with a value in the diabetic range is necessary to confirm the diagnosis.
SeeGlucose Tolerance Test
Hypoglycaemia
Glucose Tolerance Test (GTT)
Two hour 75g oral GTT
Procedure
The oral carbohydrate intake should be at least 150 g on each of the three days
immediately preceding the test. Carbohydrate restriction to less than 125 g daily may
impair glucose tolerance.
All glucose tolerance tests should be scheduled for the morning. Glucose tolerance
decreases in the afternoon and evening.
The patient must fast for 12 hours prior to the test. Water and usual medications only
are permitted throughout this period and during the test.
75 g of glucose (available in liquid form from the laboratory) should be ingested within
5–10 minutes. There is a dose adjustment for children/ adolescents of
1.75 g of glucose per kilogram of bodyweight to a maximum of 75 g. A GTT may not
be necessary in children – please contact the pathologist for advice if unsure.
1 x 5 mL fluoride oxalate whole blood is to be collected before ingestion of the glucose
load and at one hour and two hours post glucose load.
This procedure is performed by appointment at a Collection Centre.
Patient Information leaflets are available on our web site, from Stores Department on
02 6285 9813 or your nearest Collection Centre.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Oral glucose tolerance test interpretation
(75 gram load, adult, non pregnant)
1.Diabetes Mellitus
Fasting> 7.0 mmol/L and
2 hour glucose > 11.1 mmol/L
If only one value is in diabetic range, then diagnosis should be confirmed by
repeat on another day, unless there is unequivocal hyperglycaemia with acute
metabolic decompensation or obvious symptoms. The diagnosis of diabetes must
be definite.
2.Impaired Glucose Tolerance
Fasting< 7.0 mmol/L and
2 hour glucose 7.8–11.0 mmol/L
3.Impaired Fasting Glucose Tolerance
Fasting6.1–6.9 mmol/L and
2 hour glucose normal
Gestational diabetes mellitus (GDM)
GDM is a form of glucose intolerance that develops during and disappears after
parturition.
It is associated with increased fetal risk which is reduced if the GDM is treated
successfully with diet or, in the few where hyperglycaemia persists, with insulin.
A mother with GDM is at increased risk for developing diabetes in later life.
Screening test for GDM
Screening at 26–28 weeks is currently recommended for all pregnant women, though
clearly at–risk patients, e.g. previous GDM, can be evaluated earlier, even before
conception.
The test is usually done in the morning. No booking is required and the patient does
not need to fast before the tests. A suspension of 50 g of glucose is given and a single
blood collected one hour later. If the one hour glucose is equal to or above
7.8 mmol/L, the test is positive and a 2–hour 75 g GTT is ordered as the definitive test.
Diagnostic criteria for GDM (2–hour 75 g OGTT)
GDM is present if:
Fasting glucose > 5.5 mmol/l
or 2–hour glucose > 8.0mmol/
Specialized GTTs
Protocols are available for extended GTTs (3,4,5 hour GTT) to investigate
hypoglycaemia.
GTTs with insulin levels can be performed to investigate insulin resistance.
Kidson Protocol (HOMA) index GTTs can be arranged on request.
Reference:
1.Colman PG, Thomas DW, Zimmet PZ, et al. New classification and criteria for diagnosis of diabetes mellitus.
MJA 1999; 170:375–378.
2.Hoffman L, Nolan C, Wilson JD, et al. Gestational diabetes mellitus–management guidelines. The Australian
Diabetes in Pregnancy Society. MJA 1998; 169:93–97.`
G
A
Glycosuria
Glucose appears in the urine when the renal threshold has been exceeded; this
typically being at a blood glucose level around 11 mmol/L.
With advancing age, or renal impairment, the renal threshold rises so that some diabetics
have glucose–free urine even with blood glucose levels in the 12–15 mmol/L range.
Glycosuria is found in:
• Diabetes mellitus – once the renal threshold is exceeded.
• Renal glycosuria – in which the individual has a lowered renal threshold.
The GTT is entirely normal but urine specimens collected during the test show
glucose. The patient who is proven to have this uncommon condition can be
reassured that it is of no clinical significance and is unrelated to diabetes.
• In pregnancy – renal glycosuria is common in healthy non–diabetic women
during pregnancy, beginning during the first trimester and disappearing within
a week of delivery. A feature of this glycosuria is its variability from day to day,
throughout the course of the pregnancy and even during the course of a day.
Gold, heavy metal
Specimen:Urine – 24-hour urine in acid washed bottle
Plasma – EDTA tube.
Plasma should be spun and separated.
Reference Range: Supplied with report
Gonorrhoeae
Please see Neisseria gonorrhoeae.
Gram Stain
The Gram stain, described as the single most useful procedure in diagnostic
microbiology, was perfected by the Danish microbiologist Christian Gram in 1884.
When a microbiological specimen reaches the laboratory there are typically two initial
procedures:
•
•
ram stain of the specimen smeared on a slide to determine numbers of bacteria,
G
whether cocci or bacilli and whether gram–negative or gram–positive. In a case of
meningitis, for example, the gram stain gives a presumptive diagnosis and dictates
initial antibiotic treatment.
The specimen is cultured on a microbiological plate and the colonies
gram–stained next day.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Some examples:
G
A
Gram–positive organisms
cocciStaphylococcus
Streptococcus
bacilliCorynebacterium
Gram–negative organisms
cocciNeisseri
bacilli enteric bacteria:
Escherichia, Salmonella, Shigella, Enterobacter,
Klebsiella, Serratia, Proteus, Citrobacter, Yersinia
Growth Hormone
Specimen:
Serum – Gel
Reference Range: The normal values vary with age and sex
Suppression or stimulation tests are used in preference to random measurements
Elevated levels
• Pituitary gigantism
• Acromegaly
• Stress, exercise.
Low levels
• Pituitary dwarfism
• Hypopituitarism
• Glucose infusion – used in suppression tests for gigantism or acromegaly.
Gynaecomastia
Transient gynaecomastia is normal in the newborn and in adolescence and minor
degrees of persistent gynaecomastia are common in the elderly male as testosterone
levels decline with an increase in the oestrogen/testosterone ratio.
The causes of more troublesome and progressive breast enlargement include:
•
•
•
•
•
•
•
•
•
Drugs oestrogens spironolactone
Digoxincimetidine
tricyclicsdiazepam
methyldopaantiandrogens
Hypogonadism – check LH, FSH, testosterone
Liver disease
Thyrotoxicosis
Feminising tumours
Klinefelter’s syndrome.
Capital Pathology Handbook – Interpretation of Laboratory Tests
H
H
A
Haemochromatosis
Specimen:
Whole blood – EDTA
Reference Range: Supplied with report
Haemochromatosis is a condition of iron overload which is usually subclinical but
which in some patients causes systemic disease due to parenchymal tissue damage.
In its common hereditary form, the diagnosis of haemochromatosis is being made with
increasing frequency because of the ready availability of tests for ferritin, iron and iron
saturation and recently, the HFE gene.
Iron overload can also be acquired due to repeated transfusions for refractory
anaemia or to inappropriate parenteral iron over a long period of time.
Hereditary haemochromatosis (HH)
In HH the iron overload is due to excessive absorption which steadily increases the
total body mass of iron throughout life. Women are protected during child–bearing
years by menstrual loss of iron.
HH is carried on a recessive gene with the remarkably high prevalence of 1:10 in the
general population for the heterozygous (carrier) state and 1:400 for the homozygous
(affected) state.
In 1996 the HFE gene, also known as the cys282 tyr mutation, was identified as a
marker for at least 90% of HH.
Clinically HH is characterised by:
•
•
•
•
•
Lethargy, skin pigmentation, reduced sweating
Hepatic damage: elevated enzymes, cirrhosis, hepatocellular carcinoma
Endocrine damage: diabetes, hypogonadism, loss of libido, amenorrhoea,
hypopituitarism, hypothyroidism
Arthropathy
Myocardial damage: congestive cardiac failure.
Diagnosis
HH is suspected when iron saturation is consistently above 45–55% and ferritin
above 400 ug/L.
The HFE gene should be tested for and if present the test will show whether the
patient is a hetero–or homozygote.
The molecular testing currently looks for three gene mutations:
C282Y, H63D and S65C.
Haemoglobin Electrophoresis (Hb EPG)
Specimen:
Whole blood – EDTA, and Serum – Gel
Hb EPG detects abnormal and unstable haemoglobins and gives an indication
whether HbA2 or HbF are increased.
Haemoglobinopathy Screen
The screen consists of:Hb electrophoresisHbA2 quantitation
HbF quantitation
HbH body stain
Hb stability test.
Indications
•
•
•
uspected thalassaemia or haemoglobinopathy – hypochromic microcytic
S
anaemia in the absence of iron deficiency or chronic disease, particularly in
persons of Asian or Mediterranean descent.
Unexplained haemolytic anaemia.
The patient’s partner has been diagnosed with a haemoglobinopathy or
thalassaemia and the couple are in the child–bearing age group.
See
Haemoglobins, normal and abnormal
Haemoglobins, Normal and Abnormal
The haemoglobin (Hb) molecule consists of two pairs of polypeptide chains with a
haem attached to each. The porphyrias are due to disorders of haem synthesis. The
haemoglobinopathies are due to disorders in the globin chain pairs which are labelled
á â ä ã etc. In the haemoglobinopathies, the amino acid sequence in a globin chain
can be abnormal, or there can be a quantitative abnormality in globin chain production.
The main haemoglobins are:
HbA á 2â2The normal adult Hb comprising 97% of the total. Also called
HbA1, HbA1C, of interest to diabetologists, is the non–
enzymatically glycated fraction of HbA1.
HbF á 2ã2Fetal Hb comprises 70–90% of the total at birth falling to 1%
by age 2. It is increased in beta thalassaemia and hereditary
persistence of HbF (HPFH).
HbA2 á 2ä2The minor component of Hb comprising 1.5–3.2% of the total. It is
increased in beta cell thalassaemia.
HbH beta2 beta2Is increased in the α thalassaemias in which there is deletion of
1–4 of the ã chains. In the 3–chain deletion HbH disease, HbH
comprises 5–30% of total Hb.
HbSThe cause of sickle cell anaemia (HbSS) found in South Africans
and occasionally in those of Mediterranean or Middle Eastern
origin. The heterozygous stat HbAS, is asymptomatic.
SeeHaemoglobinopathy Screen
Thalassaemias
Sickle Cell Test
Capital Pathology Handbook – Interpretation of Laboratory Tests
Haemolysis
H
A
The features of haemolysis are:
•
•
•
•
Anaemia
Polychromasia–reticulocytosis
Haptoglobins–reduced or absent
Hyperbilirubinaemia.
The blood film is used to distinguish between spherocytic and non–spherocytic
haemolytic anaemia:
Spherocytes present
Spherocytes absent
Hereditary spherocytosis
Hereditary elliptocytosis
Autoimmune haemolysis
Haemoglobinopathies
(Coomb’s positive)
Enzymopathies
Drug–induced haemolysis
(e.g. G–6–PD deficiency)
Delayed transfusion reaction
PNH
ABO incompatibility in the newborn
Haemophilia
The haemophilias are due either to Factor VIII deficiency (classical haemophilia) or
Factor IX deficiency (Christmas disease). Spontaneous mutations account for 30%
of new cases in the absence of a family history. Severe haemophilia presents early in
infancy but mild haemophilia can present later in life following surgery or trauma.
Diagnosis is from:
•
•
•
•
•
Clinical bleeding history
Family history
Abnormal APTT (correcting with mixing test)
Reduced level of FVIII or FIX
DNA genotyping.
See
Coagulation Studies
Haemosiderin in Urine
A granular storage form of iron which can be seen histologically. Physiologically it is the
normal storage form of iron in bone marrow. Patholgically it is found in many parenchymal
tissues in iron–overload. S. ferritin levels reflect tissue haemosiderin load.
In chronic haemoglobinuria, haemosiderin may be detected in a fresh random urine.
Hairy Cell Leukaemia
This is a rare form of B cell leukaemia (1–2%) with a 5:1 male predominance. It is
characterised by pancytopaenia, circulating “hairy cells” and splenomegaly (80%).
Hairy cells may be infrequent in the peripheral blood, and the characteristic bone
marrow fibrosis usually leads to a dry tap at bone marrow aspiration. Treatment with
chemotherapy is highly effective, with a response and remission observed in the
majority of patients.
Haptoglobins
Specimen:
Serum – Gel
Reference Range: Supplied with report
Haptoglobin estimation is one of the screening tests for haemolysis. Hb released
by the haemolytic process combines with haptoglobin to form a complex which is
removed by the liver. It is also an acute phase reactant, migrating with the alpha 2
band on protein electrophoresis.
It is reduced by:
•
•
•
•
•
Haemolysis – haptoglobins are absent in moderate or severe haemolysis
Oestrogens, oral contraceptives, pregnancy
Acute or chronic hepatocellular liver disease
Specific genotype
Lower in first year of life.
It is elevated by:
•
•
•
Biliary obstruction
Many acute or chronic inflammatory disorders–following recovery from an infective
episode the level falls to normal within 10–14 days
Specific genotype.
HbA1c
Specimen:
Whole blood – EDTA
Reference Ranges:4.3–6.4% reference population
Clinical Evaluation
• 6.1–7.0% HbA1c indicates very good control.
• 7.1–8.0% HbA1c indicates adequate control.
• 8.1–9.0% HbA1c indicates suboptimal control.
{I Reference: ADA Position Statement. Diabetic Care 20 (Supp 1),
55–511, 1997 and RCPA–AACB Chemical Pathology QA Program, 2001}
Suggest continue monitoring diabetic control with HbA1c measurements at three
monthly intervals.
Heavy Metal Testing
Specimen:
Preferred speciman is 24-hour plain urine
Lead testing requires Whole blood – EDTA.
All other heavy metals can be done on whole blood –
Trace element tube.
It is recommended to specify individual heavy metals required.
A screen includes aluminium, arsenic, cadmium, cobalt, lead,
and mercury.
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Helicobacter pylori
Diagnosis
Histology of gastric biopsies has long been the mainstay of diagnosis of H. pylori
related disease. Serological diagnosis remains controversial, and while levels of IgG
do fall after successful eradication treatment, post–treatment monitoring can take six
months or longer to show a conclusive drop. The urea breath test is accepted as a
simple, reliable and safe diagnostic alternative.
Patient Preparation
This procedure is performed by appointment at a Collection Centre.
The patient should have nil by mouth for six hours.
The following drugs may effect the urea breath test:
Drugs to be stopped at least FOUR weeks prior to testing:
• All antibiotics (e.g. Amoxycillin, Tetracyclines, Metronidazole)
• Bismuth–containing compounds (e.g. Denol).
Drugs to be stopped at least ONE week prior to testing:
• Proton–pump inhibitors (e.g. Omerprazole, Lanzaprazole, Pantoprazole).
Patient information leaflets are available at Collection Centres or from the Client
Services Department on 02 6285 9802 or from www.capitalpath.com.au.
Hepatitis
Hepatitis is characterised by elevations of ALT and AST and a histological picture on
biopsy which may be diagnostic. When ALT and AST are above 1000 U/L with ALP
and GGT below 300 U/L, the cause will usually be viral infection. In chronic hepatitis,
changes in ALT are used as an indication of disease activity.
Main causes of hepatitis include:
• Viral infection
– Hepatitis A virus (HAV)
– Hepatitis B virus (HBV)
– Hepatitis C virus (HCV)
– EB virus (infectious mononucleosis)
Other viruses
–
HIV
–
Hepatitis D, E
–
CMV (cytomegalovirus)
–
Coxsackie virus
– Sundry other viruses causing flu–like illnesses
circulating in the community
• Autoimmune hepatitis
• Alcoholic hepatitis
• Drug associated hepatitis
• Other.
H
A
Hepatitis A Antibody (HAV)
Specimen:
Tests:
Serum – Gel
Anti–HAV IgM
Anti–HAV IgM and IgG (total antibody)
There is no test for HAV antigen.
Interpretation
The IgM antibody appears at about the time clinical symptoms develop and persists
for 6–12 months. This anti–HAV IgM is the marker for current or recent infection.
The IgG appears not long after the IgM but persists for life and is thus the marker for
HAV immune status.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Hepatitis B (HBV)
Specimen:
Serum – Gel
Tests:
HBs Ag
HBe Ag
Anti–HBs
Anti–Hbe
Anti–HBc IgM
Anti–HBc
HBV–DNA
H
A
(HBV surface antigen)
(HBV e antigen)
(antibody to HBV surface antigen)
(antibody to HB e antigen)
(IgM antibody to HBV core antigen)
(IgM and IgG antibody to HBV core)
(HBV–DNA detected by PCR)
Indications and interpretation
1.
HBs antigen
If positive indicates one of:
– Acute HBV infection (enzymes elevated) or
– Chronic HBV infection (enzymes fluctuate) or
– HBV carrier (enzymes not elevated)
2.
HBe antigen
This should be checked whenever HBs antigen is positive. If Hbe antigen (Ag) is
present, it indicates greater infectivity of the HBV infection towards sexual partner or
fetus and greater likelihood of developing chronic hepatitis. Hbe Ag is found only in
association with Hbs Ag, never on its own.
3.Anti–HBs
Indicates immunity to HBV whether naturally acquired or following immunisation. Anti–
HBs can be measured quantitatively–levels above 10 IU/L indicate immunity.
4.
Anti–HBc IgM
Is measured when current HBV infection is strongly suspected but HBs Ag and anti–
HBs are both negative. Anti–HBc IgM rises early in HBV infection and persists for
about 6 months. It fills the one month “window” between disappearance of HBs Ag and
the appearance of anti–HBs. Positive anti–HBc does not confer immunity.
5.
Anti–HBc total
Indicates past infection but unlike anti–HBs it does not indicate immunity and can
coexist with HBs Ag in carriers or chronic hepatitis. Anti–HBc remains negative after
HBV vaccination because the vaccine contains surface but not core antigen.
6.Anti–HBe
In chronic HBV infection, anti–BHe indicates low infectivity.
7.
Anti–HBV– DNA
If positive indicated HBV viraemia which is an indication for considering interferon
treatment in chronic infections.
Hepatitis C (HCV)
Specimen:
Tests:
Serum – Gel
Anti–HCV (total antibody to HCV)
HCV–RNA)
Indications and Interpretation
1.Anti–HCV
Used when hepatitis C is a possibility, e.g. IV drug users, raised liver enzymes of
unknown cause. Anti–HCV starts to rise 1–6 months after the initial infection and
persists for life. Presence of anti–HCV indicates either past infection or continuing
chronic infection.
2.HCV–RNA
This is a follow–on test when anti–HCV is positive. It is also used when clinical
suspicion of HCV infection is high but anti–HCV is negative because infection is recent
or the patient immunosuppressed.
It indicates current active HCV infection and is an indication for considering interferon
and ribavirin therapy.
Clinical course and epidemiology
HCV was identified in 1989 having previously been included in the non–A non–B
hepatitis category. A random population sample in the United States shows 1.8%
positivity.
Transmission in blood products has been largely eliminated, leaving contaminated
needles in drug users as the main source of new infections. It is the cause of 5–15%
of acute non–A, non–B hepatitis.
Mother–to–fetus transmission occurs but is infrequent, < 5%. Sexual transmission is
believed to be uncommon.
More than 50% of HCV infections become chronic with potential to develop cirrhosis
and hepatocellular carcinoma.
Treatment with a combination of interferon and ribavirin clears the virus in about 50%
of patients.
There is no vaccine yet.
Follow–up on HCV +ve patients
A positive HCV Ab test should be followed by HCV RNA test and ALT.
If HCV RNA is positive, the patient should be referred for consideration of antiviral
therapy.
An ALT > 60 units indicates active disease but a level < 60 does not exclude activity.
The patient who is HCV Ab +ve but HCV RNA –ve is presumed to be free of active
hepatitis provided the ALT is < 60. The ALT should be monitored annually and the
HCV RNA for at least 2 years to confirm it is staying negative.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Hepatitis D (HDV–Delta)
Specimen:
Tests:
Serum – Gel
Anti–Delta IgG
HDV RNA
HDV infections are found only as a co–infection or superinfection in HBV positive
patients. HDV is associated with more severe acute disease and a greater risk of
developing chronic hepatitis and its complications.
HDV infections once established, will persist for as long as HBV carriage continues,
but if HBV is eliminated HDV will go with it.
Anti–delta antibodies indicate either current active infection or past infection with immunity.
Positive HDV RNA indicates current active infection.
Hepatitis E (HEV)
Specimen:
Tests:
Serum – Gel
Anti–HEV IgG
Hepatitis E resembles hepatitis A in its faecal–oral mode of transmission. It is endemic
in India, SE Asia, China, Mexico and parts of Africa.
Presence of HEV antibodies does not distinguish current from past infection.
Identifying the virus in a small specimen of faeces may identify current infection.
Hepatitis, Autoimmune
Specimen:
Serum – Gel
Immunoglobulins
Antimitochondrial antibodies
Tests:
Liver/kidney microsomal antibodies
ANA
Smooth muscle antibodies
Anti–ds DNA
Autoimmune hepatitis, formerly known as chronic active hepatitis and before that lupoid
hepatitis, affects predominantly young women. The hepatitis can be of all grades of
severity ranging from minor enzyme elevations in a well person to a clinical laboratory
picture like that of severe viral hepatitis. Diagnosis is by excluding viral causes; by
finding specific autoantibodies; by markedly elevated IgG; and by liver biopsy.
Smooth muscle antibodies (SMA) in high concentration (> 80) define autoimmune
hepatitis.
Antimitochondrial antibodies (AMA) and total IgM may be elevated but these are
typically the markers of the other autoimmune liver disease, primary biliary cirrhosis.
Liver/kidney microsomal antibodies (LKM) are uncommon but help in disease
classification. Titres tend to be lower in viral hepatitis than in autoimmune hepatitis.
H
A
Hereditary Spherocytosis
Specimen:
Whole blood – EDTA
A relatively common haemolytic disorder found in all races and typically inherited as
an autosomal dominant. In the 25% of individuals who have unaffected parents, the
disease is assumed to be due to autosomal recessive inheritance or a spontaneous
mutation.
More severe forms present in the first 10 years of life but milder forms may be
discovered incidentally either on a blood count or because of a minimally elevated
bilirubin resembling Gilbert’s syndrome.
Laboratory tests typically show the features of a haemolytic disorder – increased
reticulocytes, reduced haptoglobins, hyperbilirubinaemia, anaemia.
The more specific findings are:
•
•
Spherocytes on the blood film
Increased osmotic fragility of red cells.
Herpes Simplex Virus (HSV)
PCR (preferred)
Specimen:Fluid or vesicle scrape
Collected in a sterile container, a viral swab, or plain swab.
Immunofluorescence and culture is being superceded by PCR.
There are two types of herpes simplex, HSV1 and HSV2.
HSV1 commonly gives rise to recurrent oral infections (“cold sores”) and HSV2 to
recurrent genital infections but either can be found at either site.
Herpes Type 6 (Roseola infantum)
Specimen:
Serum – Gel
Reference Range: Supplied with report
HIAA (5–Hydroxy Indole Acetic Acid)
Specimen:
24-hour urine (HCL preservative)
Reference Range: Supplied with report
Avoid the following drugs and foods for 3 days before collecting the urine:
• Phenothiazines
• Methyldopa
• Naproxen
• Cough mixture
• Acetaminophen
• Bananas
• Plums
• Pineapple
• Fruit juices
• Kiwifruit
• Tomatoes
• Eggplant
• Avocado
• Walnuts
• Pecans.
In carcinoid syndrome the HIAA is usually elevated. Carcinoid syndrome, due
to release of serotonin from a carcinoid tumour, is characterised by cutaneous
flushing, diarrhoea, valvular heart disease, and less often, by wheezing, paroxysmal
Capital Pathology Handbook – Interpretation of Laboratory Tests
hypotension and telangiectasia. HIAA is the breakdown product of serotonin and is
usually measured in response to a history of flushing though this is more commonly
due to menopause, alcohol (particularly in combination with a sulphonylurea or
disulfiram). Phaeochromocytoma is infrequently a cause of flushing.
Histopathology
Specimens for routine histopathology should be placed promptly in formalin fixative
(10% buffered formalin) after surgical removal.
Tissue specimen containers with fixative added are suitable for most biopsies and
small excision specimens. Other containers are available for larger specimens. Ideally,
the container should hold a volume of fixative at least 10 times that of the specimen.
Tissue for the following investigations should not be placed in formalin:
•
•
•
•
•
Frozen section
Liver iron estimation
Microbiological investigation
Flow cytometry
Disaccharidase assay on
small bowel biopsy
•
•
•
•
Immunofluorescence
Chromosome studies
Crystal identification
Testicular biopsies for infertility.
Crystal Identification in Tissue Specimens
In cases of suspected gout where the detection and identification of crystals is
required in tissue specimens, special collection procedures should be followed.
The tissue specimen should be placed into a specially marked “specimen container”
containing absolute alcohol. Where routine histopathology is required in addition to
crystal detection, the specimen should be divided. Place one portion in formalin (as
normal), the other into alcohol.
Appropriate specimen collection jars containing alcohol are available from the
Histology Department by phoning 02 6285 9855.
Disaccharidase Assays
These tests are performed on small bowel biopsies of patients with pain or diarrhoea
where sugar intolerance is suspected. Wrap the biopsy loosely in parafilm and then in
foil, store in a small screw cap vial and immediately freeze. Phone Couriers on
02 6285 9877 for rapid pick up of specimen.
Electron Microscopy
With the continuing advancements in immunohistochemistry, electron microscopy is
less commonly required as an adjunct to light microscopy.
Applications of this technique include the further identification of undifferentiated
malignancies, interpretation of renal biopsies and identification of some microbial
organisms.
Tissue for electron microscopy should be finely sliced and placed in glutaraldehyde.
Please phone the Histology Department on 02 6285 9855 for further details.
H
A
Frozen Sections
The histopathologists are available by arrangement for frozen section diagnosis. For
frozen section bookings, please phone the Histology Department on 02 6285 9857.
It is preferred that at least 24 hours notice is given to facilitate arrangements for a
pathologist and technologist to be on site for the operation.
Cryostat facilities are available at Calvary John James Hospital, Calvary Hospital and
The National Capital Private Hospital to provide an on–site frozen section diagnosis.
There is also a cryostat at the laboratory.
Hormone Receptor Assays
Tumour hormone receptor analysis (usually breast carcinoma) is now performed on
formalin fixed tissue by the immunoperoxidase method, and no special collection
procedure is necessary.
Liver Iron Content
Liver biopsies for iron estimation require special collection procedures and should not
be placed in fixative.
Please phone the Histology Department on 02 6285 9855 for further information.
Lymph Node Biopsy
Lymph nodes can be surgically excised or sampled via fine needle aspiration
for microbiological examination, suspected primary or metastatic neoplasia,
lymphoproliferative diseases and other causes of lymphadenopathy. Lymph node
biopsy may also be useful for staging carcinoma and lymphoma, particularly if
lymphoproliferative diseases are suspected. For optimal results, the node should be
removed with minimal trauma and submitted fresh to the laboratory in a clean dry
specimen container that is kept cool, (but not frozen).
In the laboratory the tissue may be examined using frozen section, smears, imprints,
and flow cytometry while fixed tissue may be examined using light microscopy and
immunohistochemistry.
Microbiological Investigation of Tissue Specimens
Tissue specimens for culture should be obtained using full aseptic technique and
transferred to an appropriate container without contamination. Transport should be in
a “specimen container” with the addition of a small amount of sterile saline to prevent
drying out of the tissue. Specimens must not be transported in formalin.
Microsatellite Instability for Hereditary Colorectal Cancer
An immunoperoxidase antibody staining method is used to examine for the gene
products. This is performed on routinely processed tissue.
Skin Biopsy for Inflammatory Skin Diseases
A detailed clinical history, including medications, clinical appearance and distribution
of the skin lesions can be very helpful in arriving at an accurate diagnosis. Fresh
tissue from skin biopsies can be examined by immunofluorescence and, if indicated,
microbiology.
Capital Pathology Handbook – Interpretation of Laboratory Tests
For the investigation of skin lesions such as bullous rashes and other dermatoses
two punch or incisional biopsies are recommended. One should be placed in buffered
10% formalin for light microscopy and the other placed in transport medium for
immunofluorescence (see skin biopsy).
Please phone the Histology Department on 02 6285 9855 for appropriate transport
medium.
Testicular Biopsy for Infertility
Formalin is not an appropriate fixative for preservation and interpretation of testicular
tissue. Bouin’s fixative is used for this purpose.
Please phone the Histology Department on 02 6285 9867 for Bouin’s fixative and for
further instructions.
HIV (Human Immunodeficiency Virus)
Specimen:
Serum – Gel
HIV Antigen / Antibody
At Capital Pathology routine initial serology provides testing for the p24 antigen
and antibodies to HIV-1 and HIV-2. The antigen may be present early in infection.
The antibodies can be seen with recent or past exposure. Negative results do not
exclude infection. Repeat serology may be required, for instance if serum has been
taken only a short time after exposure. Reactivity on the initial diagnostic test will
result in further confirmatory tests being required, which are referred to a specialized
reference laboratory. Supplementary HIV tests may include HIV viral load testing, HIV
by Western Blot and HIV by PCR. Although the results from the routine initial tests
are readily available, results from supplementary testing may take longer. Medicare
benefit rebate is now available for HIV serology testing.
HIV PCR (Viral Load)–Quantitative
Specimen:
Whole blood – EDTA x 2
Reference Range: Supplied with report
H
A
HLA–B27 Antigen (Human Leucocyte Antigen B27)
Specimen:Whole blood – EDTA
Keep specimen at room temperature.
Collect Monday–Friday (Friday am only).
•
•
•
•
•
•
HLA B27 has a frequency of 7–10% in the general population but a much higher
frequency in the seronegative spondyloarthropathies which are characterised by:
Sacroileitis and spondylitis, causing low back pain and loss of lumbar mobility
Tendonitis, including heel pain and other pain at muscle and tendinous insertion
Oligoarthritis
Urethritis
Iritis and conjunctivitis.
As indicated by the term seronegative, all of these disorders are negative for
rheumatoid factor.
The principal members of the group with the % positivity for HLA B27 are:
•
•
•
•
•
Ankylosing spondylitis (95%– 98%)
Reiter’s disease (90%)
Arthropathies associated with ulcerative colitis and Crohn’s disease (70%)
Post–enteric and post–venereal reactive arthritis (70%)
Psoriatic arthritis (30%).
HLA Tissue Typing
Contact laboratory prior to collection.
Specimen:Whole blood ACD, EDTA, plain clot tube
The number and type of gel tubes for collection depends on the
indication and history.
Specimen collection is available at all the Collection Centres.
Red Cross Sydney provides further information on specific
requirements.
Can be collected Mondays to Thursdays, however a booking
may need to be made with Red Cross by the doctor or Collection
Centre prior to specimen collection.
Red Cross 02 9234 2322.
For further clinical information please contact the Director of
Clinical Pathology on 02 6285 9895.
Homocysteine
Specimen:Plasma (Lithium heparin-gel tube)
Spin as soon as possible. Patient should be fasting.
Reference Range: Supplied with report
Previously, clinical interest in homocysteine in blood was confined to the rare inborn
error of metabolism homocystinuria in which plasma homocysteine is markedly
elevated to above 100 umol/L.
More recently an association has been described between mild to moderate
hyperhomocysteinaemia (15–100 umol/L) and cardiovascular disease and many now regard
it as an independent risk factor for coronary, cerebral and peripheral arterial disease.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Increasing intake of folic acid and to a lesser extent vitamin B6 and perhaps B12 may
lower homocysteine levels. A daily supplement of 400 ug folic acid will in most patients
increase serum folate above 15 nmol/L and drop plasma homocysteine to a low
plateau. As yet there is no proof of a favourable effect on cardiovascular outcomes.
Homocysteine elevation above 20 umol/L is also a risk factor for venous thrombosis
(2–3 x increase).
Hookworm
Hookworm refers to intestinal infestation with Ancylostoma duodenale (found in
the Mediterranean basin, the Middle East, India and China) or Necator americanus
(found in the Americas, sub–Saharan Africa and SE Asia). Heavy infestations are an
important cause of iron–deficiency anaemia.
Diagnosis is by identifying eggs in faeces.
Treatment is with mebendazole 100 mg, 12–hourly for 3 days.
Hormone Assessment (Female)
Evaluation of Female Infertility
Suggested
scheme
for
the work-up
Female Infertility
Suggested
scheme
for the
work-up
of Femaleof
Infertility
PRL: Elevated
Hyperprolactinaemia
FSH: Elevated
LH: Elevated
Ovarian failure
FSH: Normal
LH: Elevated
Polycystic ovaries
Elevated
Ovulation confirmed
Normal
Ovulation not confirmed
Progesterone
ale Infertility Ix
Serum: FSH, LH, Progesterone (day 21), Prolactin (PRL)
Female 1
Tubal disease
Endometriosis
Male infertility
Exclude:
H
A
Female 2
Irregular Periods Ix
Investigative protocols
Evaluation of irregular periods
Suggested scheme for evaluation of a patient
Evaluation
of irregular periods
with irregular periods
History/
Examination
Serum: FSH, LH, Prolactin (PRL)
PRL: Elevated
Hyperprolactinaemia
FSH: Elevated
LH: Elevated
Ovarian failure
? Menopause
FSH: Normal
LH: Elevated
? Polycystic ovaries
? Ovulation defect
FSH: Normal
LH: Normal
Routine gynaecological
investigations
FSH: Low
LH: Low
? Hypothalmic/
Pituitary disease
? Oral contraceptives
? Weight loss/dieting
? Excessive exercise
Capital Pathology Handbook – Interpretation of Laboratory Tests
Hormonal changes during menstrual cycle
Hormonal changes during menstrual cycle
50
LH (U/L)
Menopause:
10–60U/L
Female 4
40
Ovulatory phase
30
20
Menopause q
10
0
15
2
4
6 8 10 12 14 16 18 20 22 24 26 28
Menses
Days of cycle
FSH (U/L)
Menopause:
20–150U/L
10
Menstrual Hormone Graphs
5
0
1,500
4
6 8 10 12 14 16 18 20 22 24 26 28
Menses
Days of cycle
Oestradiol (pmol/L)
Menopause:
40–180pmol/L
1,250
1,000
750
500
250
Menopause t
0
Investigative protocols
2
100
2
4
6 8 10 12 14 16 18 20 22 24 26 28
Menses
Days of cycle
Progesterone (nmol/L)
Menopause:
< 3.0nmol/L
Ovulation:
> 20nmol/L
80
60
40
Ovulation q
20
0
2
4
6 8 10 12 14 16 18 20 22 24 26 28
Menses
Days of cycle
Follicular phase
108 Sullivan Nicolaides Pathology
Luteal phase
H
A
Evaluation of ? Androgen excess
and/or Hirsutism (Female)
Suggested scheme for evaluation of Hirsuitism/
Evaluation
of ?Androgen
Hyperandrogenism
in women excess and/or Hirsutism (Female)
Female 5
Serum: DHEAS 1 ; Testosterone; SHBG 2
DHEAS
Normal
Elevated
Exclude
adrenal
disease
Androgen overproduction unlikely
< 55
Testosterone : SHBG ratio
> 55
Evidence of androgen excess (adrenal/ovary)
Testosterone
Normal
Consider:
Polycystic ovaries
Testosterone
administration
Congenital adrenal
hyperplasia
Elevated
Consider:
Androgen-producing
tumour (ovary or
adrenal)
Testosterone
administration
1. DHEAS: dehydroepiandrosterone sulfate
2. SHBG: sex hormone binding globulin
Investigative protocols ? Androgen Excess (Female) Ix
Unlikely to
have adrenal
pathology
Investigative protocols
109
Capital Pathology Handbook – Interpretation of Laboratory Tests
Human Papillomavirus (HPV)
Capital Pathology offers DNA testing for high–risk strains of Human papillomavirus
(HPV) using the Cobas 4800 HPV DNA assay.
HPV infection is now considered a necessary but insufficient cause of cervical cancer.
Co–factors are required to transform the cells into truly neoplastic cells. There are,
however, many different sub–types of HPV and only some of these have been shown
to be associated with the development of high–grade cervical lesions. The HPV test
we offer identifies only high risk subtypes.
The HPV test is designed to be used in conjunction with the Pap smear result to assist
doctors in making decisions on the management and follow–up of selected patients.
The basis of the Test?
• The test used is the Cobas 4800 HPV DNA assay. This is more specific and
sensitive than previous tests.
• The test specifically identifies HPV 16 and HPV 18, the HPV genotypes most
strongly associated with cervical neoplasia. A further 12 high risk genotypes
(31,33,35,39,45,51,52,56,58,59,66 and 68) are tested and reported as a group
but not individually identified. Infection with these genotypes confers a lower risk
of developing cervical cancer than infection with HPV 16 or HPV 18.
• The result is reported as detected or not detected.
When is it indicated?
The clinical usefulness of this test is still being evaluated in many trials. However,
the test may be of benefit in the following situations:
Management of smears showing persistent low grade non–specific changes
HPV testing may clarify management options in smears with mild atypia or uncertain
changes. A positive HPV test indicates that a high grade intraepithelial lesion may
be present. A negative HPV test may allow follow–up with further smears. At present
there is no Medicare rebate for HPV testing in this circumstance.
Management of low grade (CIN 1) lesions
Neither cytology nor histological biopsy can predict which low–grade lesions will
progress. Many CIN1 lesions will harbour a high risk HPV subtype, but many of these
will regress. HPV testing in this circumstance will not predict which lesions will regress,
persist or progress. HPV testing in this situation is not indicated.
Follow–up after treatment
Persistence of high risk HPV after treatment carries a risk of progression. Testing for
HPV after treatment is therefore an effective way of monitoring cure. This is the only
situation where there is a Medicare rebate for HPV testing. This is the “test of cure”
following treatment of a high grade squamous intraepithelial lesion (see below).
Management of equivocal histology
A negative HPV test and a normal colposcopy give a negative predictive value of
98%. Testing for HPV therefore can assist management decisions when a negative or
equivocal result is reported on a colposcopically directed biopsy.
H
A
How is it collected?
The HPV test can be collected:
•
•
•
Co-collection of the conventional smear and the ThinPrep pap test –
the conventional Pap smear is performed and the instrument is then rinsed in
the ThinPrep vial. The HPV test can be performed from the sediment remaining
in the vial after the ThinPrep test has been completed. There is no need to take
a separate HPV sample.
As a separate specimen using a cervex sampler and transferring the material into
the ThinPrep vial.
At colposcopy – the HPV test is collected prior to the application of acetic acid or
any other solution.
Cost
The HPV test has no associated Medicare rebate except for follow up of high grade
lesions after treatment. Currently the cost of HPV testing is $70 if The Medicare
criteria are not met.
Management of women with treated high grade squamous intraepithelial lesions:
There is now a Medicare rebate for HPV testing in the following circumstance alone.
When a woman has received excisional or ablative treatment for a high grade
squamous intraepithelial lesion (HSIL) of the cervix, or who within the last 2 years has
had a positive HPV test after exisional or ablative treatment for high grade squamous
intraepithelial lesions of the cervix, or who is already undergoing annual cytology
review for follow up of a previously treated HSIL, then this test can be ordered
to a maximum of two requests per 24 month period. When there have been two
consecutive negative Pap smears and HPV tests, then the woman can return to the
normal screening interval. Therefore if ordering HPV testing for these circumstances,
please clearly state the reason for the HPV test on the pathology request form. We will
then be able to bill for the correct item which will be rebatable from Medicare. If a
patient does not fulfil the criteria as per the Medicare Schedule, then a non rebatable
account will be issued to the patient for the cost of the HPV DNA assay.
See
Cervical Cytology
Hydatid Serology
Specimen:
Serum – Gel
Reference Range: Supplied with report
Hydroxyproline
Specimen:Fast overnight. In the morning, void all the urine and
discard. Do not have any breakfast (water only).
After 2 hours collect 10 mL urine in specimen container.
Keep cool. May be refrigerated for up to one week,
otherwise freeze.
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
5–Hydroxytryptamine (5HT) (Serotonin)
Specimen:Serum – Gel tube
Spin, separate and freeze serum or
24–hour urine (HCL preservative).
Patient should follow special diet. Please contact your nearest
Collection Centre or the Client Services Department on
02 6285 9802 for information sheet.
Reference Range: Supplied with report
See
Carcinoid Syndrome
17–Hydroxy Progesterone (17–OHP)
Specimen:
Serum – Gel
Reference Range: Supplied with report
17–OHP has replaced urinary pregnanetriol as the best screening test for the most
common form of congenital adrenal hyperplasia (CAH).
17–OHP is a precursor of cortisol, androgens and oestrogens. In CAH, a block in the
cortisol pathway leads to overproduction of 17–OHP and androgens.
See
Congenital Adrenal Hyperplasia
Hypertension–Endocrine Causes
These include:
•
•
•
•
•
•
Conn’s syndrome
Cushing’s syndrome
Phaeochromocytoma
Renal artery stenosis
Acromegaly
Hyperthyroidism.
H
A
Hypogammaglobulinaemia
Can be found in:
•
•
•
•
•
•
Myeloma
Chronic lymphocytic leukaemia
Lymphoma
Non–selective protein loss
Transiently with a recent viral infection
Congenital deficiency.
See
Immunoglobulins IgA, IgG, IgM
Hypoglycaemia
The glucose threshold at which symptoms of hypoglycaemia develop varies widely
between individuals but in general a glucose level < 3.0–3.5 mmol/L can be described
as hypoglycaemic except in neonates where the lower limit of “normal” is about
2.5 mmol/L.
Hypoglycaemia in diabetics treated with insulin or sulphonylureas
Symptoms are common, diverse and often misinterpreted or overlooked. The transient
nature of the hypoglycaemia means that laboratory collects usually miss the trough
but any value below 4.0 is suspicious. Symptoms and signs include irritability,
tremulousness, faintness, sweating, pallor, hunger, weight gain, headaches (headache
on waking may be due to nocturnal hypoglycaemia), personality change, deterioration
in level of consciousness, coma.
Reactive (functional) hypoglycaemia
This common but elusive syndrome with its symptoms of irritability, tremulousness
and hunger (“faint with hunger”) is not easily diagnosed by the laboratory. It is seen
in children, young adults, and to a lesser extent older people. The symptoms are felt
some 2–5 hours after food, not while fasting.
The extended GTT has been the traditional test but is a burden to the patient and has
too many false positives and false negatives.
A marginally more useful test is the measured blood glucose during an attack but the
logistics of this are often not easy.
The best test is a trial diet which, if successful, combines diagnosis with therapy.
Sweet refined carbohydrates are changed to unrefined carbohydrates with one or two
snacks added between meals.
Fasting hypoglycaemias
These are much less common than the non–fasting varieties and much more likely to
have an identifiable organic basis.
The first test is to measure a fasting glucose. If low it should be followed by a fasting
insulin and repeat glucose.
•
aised or inappropriately normal fasting insulin
R
– Insulinoma
– Exogenous insulin or sulphonylureas.
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
ow fasting insulin
L
– Advanced liver disease
– malignancy
– insulin autoantibodies
– alcohol
– pituitary/adrenal insufficiency.
Neonatal hypoglycaemia
The at–risk infants are:
•
•
Low birth weight
Infants of diabetic mothers.
In both, blood glucose levels should be monitored, beginning an hour after birth,
and treatment given if levels are below 2.0–2.5 mmol/L. Glucose levels can rise and
fall quickly during the neonatal period. Clinical signs are an inadequate indication of
hypoglycaemia.
Hypopituitarism
Usually due to a tumour of the pituitary or hypothalamus but pituitary infarction or
haemorrhage can occur in a normal pituitary, classically as Sheehan’s syndrome
after obstetric haemorrhage, but also in other situations, including bleeding disorders,
cardiac by–pass surgery and some imaging procedures.
Clinical hypopituitarism may present as amenorrhoea, hypogonadism, hypothyroidism,
adrenal insufficiency, hyponatraemia.
Useful tests include:
Thyroid T4 low but TSH usually normal. The TSH measured in this situation can
coexist with clinical hypothyroidism so is assumed to be biologically inert.
Gonads FSH and LH may be low, particularly in post–menopausal women who
otherwise have high levels. Testosterone may be low in men.
Hyposplenism
After splenectomy, red cells show Howell–Jolly bodies, targeting and spherocytosis
and there is thrombocytosis and leukocytosis which may or may not persist.
A hyposplenic blood picture can also be seen in other conditions associated with
splenic atrophy:
•
•
•
•
Coeliac disease
Inflammatory bowel disease
Autoimmune disease, particularly with haemolysis
Sickle cell disease.
After splenectomy a patient may have increased susceptibility to infection which can
be overwhelming in children. Pneumococci are the usual cause but other organisms
such as Haemophilus or meningoccus may be responsible. Pneumococcal and
meningococcal vaccines can be given every 5–7 years or penicillin prophylaxis in
susceptible patients.
H
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
I
I
A
Idiopathic Immune Thrombocytopenic Purpura (ITP)
A relatively common autoimmune disorder characterised by an isolated
thrombocytopenia in the absence of drugs, chemicals or any haematological or non–
haematological disorders.
Acute immune thrombocytopenia is distinguished from acute viral thrombocytopenia
by the clinical course with the latter usually recovering within a few days without the
requirement for specific therapy. Chronic immune thrombocytopenia is diagnosed
when the thrombocytopenia persists for more than 6 months. It may have a viral
trigger, occur as part of an autoimmune disorder (e.g. SLE), arise in association with a
lymphoproliferative disease or as an idiopathic finding.
ITP is usually a diagnosis of exclusion. Appropriate investigations may include a
coagulation screen with bleeding time (to evaluate a possible associated platelet
dysfunction), autoimmune investigations (ANA, anti–DNA rheumatoid factor,
anticardiolipin antibodies, platelet antibodies), lupus anticoagulant screen, protein
electrophoresis and immunoglobulins, HIV.
Treatment is either with prednisone or intravenous immunoglobulin, particularly when
the platelets fall below 20x109/L.
SeeThrombocytopenia
IgA Deficiency
A selective reduction in IgA is a common congenital immune deficiency syndrome
occurring in 1:700 people. Most individuals are asymptomatic. IgA deficiency is
associated with autoimmune disorders including coeliac disease, SLE, autoimmune
haemolytic anaemia, rheumatoid arthritis and thyroiditis.
Anaphylaxis may occur as a result of IgG antibodies to IgA when the individual is
exposed to blood transfusion or gammaglobulin therapy.
See
Immunoglobulins IgA, IgG, IgM
IgE total
Specimen:
Serum – Gel
Reference Range: Supplied with report
IgE antibodies mediate Type I allergic reactions – allergic rhinitis, asthma, atopic
dermatitis, anaphylaxis.
Total IgE is elevated in patients with multiple allergies and parasitic infestations.
Very high levels, above 1000 IU/L, are found with atopic dermatitis, fungal sinusitis
and allergic pulmonary aspergillosis.
SeeRAST
Imipramine
Specimen:
Plasma – Lithium heparin
Reference Range: Supplied with report
See
Antidepressant Drugs, tricyclic
Immunoglobulin G Subclasses
The four IgG subclasses are numbered 1 to 4. In some patients, recurrent respiratory
tract infections are associated with Ig subclass deficiency even though the total IgG is
within the usual range.
Immunoglobulins, IgA, IgG, IgM
Specimen:
Serum – Gel
Reference Range: Supplied with report
Quantitation of immunoglobulins, particularly paraproteins, is method–dependent so
when monitoring a patient with myeloma or an apparently benign paraprotein, the
immunoglobulins will ideally be measured by the same laboratory using the same
method. In practice, laboratories change methods at times so when a result shows an
unexpected rise or fall, the laboratory should be consulted.
Immunoglobulins are antibodies and constitute the gamma fraction of serum globulins.
Abnormalities of immunoglobulins include:
•
•
•
Monoclonal increases, benign or malignant
Polyclonal increases as seen in chronic inflammatory disorders
Polyclonal decreases: immune paresis.
Influenza
Specimens: Swab – One dry Nasopharangeal or viral throat swab for
Influenza viral PCR.
Serology – Serum – Gel
The accompanying request form is to request Influenza PCR .
If serology is required – serum ( gel tube ) 5–10mls of blood.
The diagnosis of suspected influenza in a patient with respiratory symptoms can be
made by direct viral antigen detection (preferred) or by indirect antibody detection.
The preferred test is influenza A and B viral detection by polymerase chain reaction
(PCR) which is highly sensitive and specific. There is a rapid immunochromatographic
test for influenza viral detection, however it has limited sensitivity and specificity and
should be used only as a guide. Serology for influenza specific antibodies is available,
and a positive diagnosis is defined by a single high titre or by a rise in titre by at least
four fold in repeat serum specimens 10–14 days apart. Serology is less specific and
may take longer for results.
If Avian influenza is suspected on the basis of the history and clinical findings, contact
the local area public health unit first for advice on how to proceed. Depending on
the presentation the patient may need to be triaged via the public health unit, with
specimens and tests referred to a nominated laboratory with access to PCR tests for
Avian influenza.
Capital Pathology Handbook – Interpretation of Laboratory Tests
INR (International Normalised Ratio)
Specimen:
Plasma – Sodium Citrate
The INR is the test used to monitor warfarin anticoagulation therapy which is being
increasingly used across the range of thromboembolic disease. Warfarin is a vitamin
K antagonist and acts by reducing levels of Factors II (prothrombin), VII, IX, and X.
The INR is a standardised prothrombin ratio calibrated so that INR results from one
laboratory are directly comparable with those from another.
Recommended therapeutic range: clinical state
1.5–2.0
Consider in cases where risk of cerebral haemorrhage is high, i.e.
over 75yrs, for some indications such as atrial fibrillation (AF). Long–
term prevention of CVT/PE after > 3 months after last event.
2.0–3.0
Prevention of DVT, therapy for DVT or PE, preventing systemic
embolism (AF, valvular heart disease, after AMI, tissue heart
valves–1st three months).
2.5–3.5
Bileaflet mechanical heart valve (aortic).
3.0–4.5
Mechanical prosthetic heart valve (high risk), secondary prophylaxis
in antiphospholipid syndrome.
Reference:
ACT Regional Guidelines for the Use of Warfarin, Sept 2003
Insulin
Specimen:Serum – Gel
Patient should be fasting.
Reference Range: Fasting insulin < 12mU/L makes insulin resistance unlikely
Insulin measurements may be requested in conjunction with a Glucose Tolerance Test.
Insulin-like Growth Hormone Factor–1 (IGF–1)
Specimen:
Serum – Gel
Reference Range: Supplied with report
The action of Human Growth Hormone (HGH) from the pituitary is mediated through
IGF–1 which is formed in the liver. As well as its effects on growth, it has insulin–like
actions. IGF–1 synthesis is suppressed by malnutrition as well as HGH deficiency.
IGF–1 and HGH are measured in suspected acromegaly and in pituitary dwarfism.
I
A
Intrinsic Factor Antibodies
Specimen:
Serum – Gel
Reference Range: Not detected
SeeVitamin B12
Pernicious Anaemia
Iron
Specimen:
Serum – Gel
Reference Range: Child 4.5–27 umol/L
Adult 9–27 umol/L
Serum iron levels show a marked diurnal variation with morning levels higher by 30%.
They are also rapidly depressed to low levels by acute or chronic infections or any
other chronic disorder.
Because of these large fluctuations, serum iron is of little diagnostic use on its own
but when combined with iron–binding capacity/transferrin and ferritin, total body iron
deficiency or excess can usually be defined.
Serum iron is elevated by:
•
•
•
•
•
•
Oral or parenteral iron supplements
Haemochromatosis
Alcohol
Hepatitis
Oestrogens/oral contraceptives
Iron poisoning – children absorb oral iron easily and in acute poisoning can
present with levels in the 100–500 umol/L range.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Iron deficiency
I
A
Useful tests
Ferritin
The most sensitive and specific test. If low it is almost pathognomonic
of iron deficiency, latent or overt, but a normal result does not exclude
iron deficiency as coexisting inflammatory disease can elevate ferritin
independently.
MCV & MCH
Reduced, but not until ferritin is < 10
Hb
Reduced, but anaemia develops later than the other changes
Blood film
Oval cells, hypochromia, microcytosis
Iron
Reduced, but many other conditions cause a fall in serum iron
IBC
Increased (may be normal if inflammation coexists)
% saturation
Reduced
Aetiology of iron deficiency
The treatment of iron deficiency must always be accompanied by a search for the cause.
Deficiency is so common in women of child–bearing age that it is easy to attribute all
deficiencies in women to menstrual loss while forgetting this in not possible after the
menopause – nor in males of any age. Deficiencies fall into three broad groups:
1. Chronic blood loss
Female genital tract:
• Menstrual loss
• Pregnancies
• Tumours of female genital tract with abnormal bleeding.
Gastrointestinal loss:
• Oesophagus, stomach, duodenum–peptic ulcer, NSAIDs, hiatus hernia, varices
• Large bowel – tumours (iron deficiency is a common presentation), ulcerative
colitis, haemorrhoids
• Small bowel – hookworm, congenital vascular malformations, Meckel’s
diverticulum.
Other:
• Epistaxis, haematuria, haemoptysis, telangiectasia.
2. Dietary insufficiency
Mainly in infants, adolescents and the elderly.
3. Malabsorption
Coeliac disease particularly.
Iron deficiency in athletes
Iron deficiency is more common in athletes for three reasons:
•
•
•
They may be on relatively low iron diets
They frequently suffer occult gastrointestinal blood loss
They may suffer haemoglobinuria with urinary iron loss.
For these reasons many athletes take oral iron supplements when their ferritins are
towards the lower end of the reference range.
Islet Cell Antibodies (ICA)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
J
J
A
Japanese Encephalitis Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Jaundice
Serum Bilirubin
<40µmol/L
?Carotenaemia
Dietary
Myxocedema
Both Normal
Isolated
hyperbilirubinaemia
> 40µmol/L
Serum ALT and ALP
ALT (U/L)
< 200
< 200
? Not Liver Disease
? Mixed
> 200
Predominant
Cholestatic
ALP (U/L)
Predominant
hepatocellular
< 250
200–400
ALP (U/L)
250–350
> 350
> 400
Predominant
hepatocellular or
mixed
Predominant
Cholestatic or
mixed
< 250
Predominant
hepatocellular
> 250
Mixed
ALP (U/L)
Joint Aspirate
See
Synovial Aspirate
Capital Pathology Handbook – Interpretation of Laboratory Tests
K
K
A
Ketones
Specimen:
Random urine
This test, which is part of routine urinalysis, is usually done by a dipstick method.
Mild positives occur with starvation or illness. Moderate or strong positives in Type I
diabetics indicate poor control requiring additional insulin and rehydration – urgently if
there is significant acidosis.
“Ketostix” detects acetoacetate which accounts for only a small percentage of “ketone
bodies”, the greater portion being beta–hydroxybutyrate.
Klebsiella pneumoniae
An aerobic gram–negative enteric bacillus. Found on mucosal surfaces and in the
faeces of about 5% of healthy people. It occasionally causes pneumonia as well as
biliary and urinary tract infections.
Kleihauer Stain for Fetal Haemoglobin
Specimen:
Maternal whole blood – EDTA
Reference Range: Supplied with report
Klinefelter Syndrome
XXY is the usual karyotype. Testosterone levels are typically reduced and FSH and LH
increased. Other pituitary tests are usually normal.
Clinically, patients are often unusually tall with long legs. Testes are small, pubic hair
scant and there may be gynaecomastia.
Capital Pathology Handbook – Interpretation of Laboratory Tests
L
L
A
Lactate Dehydrogenase (LD) (LDH)
Specimen:
Serum – Gel
Reference Range: Adults 100–250 U/L
LDH is an enzyme of low specificity found in liver, myocardium, skeletal muscle and
red cells. It is also found in some malignancies, notably lymphomas and particularly
non–Hodgkin’s lymphoma.
Elevations can be due to:
•
•
•
•
•
•
•
•
ymphomas, up to 2–3x the upper limit of the reference range – the level gives an
L
estimate of tumour bulk
Other tumours, especially germ cell
Myocardial infarction, increase commences 12–24 hours after infarction,
disappearing after 7–12 days
Megaloblastic anaemias due to B12 or folate deficiency
Haemolytic anaemia
Artefactual haemolysis due to faulty specimen or collection
Liver disease, particularly hepatitis
Skeletal muscle damage.
Of the LD isoenzymes, LDI is derived from myocardium or erythrocytes, LD5 from liver.
Lactate Dehydrogenase (LDH) Interpretations
Elevated LDH
Lead, blood
Specimen:
Whole blood – EDTA
Reference Range: Supplied with report
Domestic exposure occurs when old lead–based paints are being sanded or burnt off.
Children are at risk when they ingest paint fragments or dust, as are pets when they
lick their fur.
High levels should be repeated in 3–4 weeks.
Lead poisoning interferes with porphyrin metabolism causing elevation of urinary
coproporphyrin, PBG and ALA. The porphyrin changes are a better indicator of toxicity
than the blood lead on its own but are not used for routine industrial testing. Red cells
may show basophilic stippling.
Lead, urine
Specimen:
24-hour urine (nil preservative)
Reference Range: Supplied with report
Urine leads are used for monitoring chelation therapy in lead poisoning after the
diagnosis has been established using blood lead levels.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Legionella Antibodies
Specimen:Serum – Gel
Sputum for culture.
There are more than 30 species of Legionella, the cause of Legionnaire’s disease.
The organism is found in hot water systems or the humidified air of air–conditioning
systems, causing both sporadic cases and institutional outbreaks of atypical
pneumonia.
Culture is the diagnostic method of choice. Legionella culture must be asked
for specifically as it requires the use of specially prepared media. Specimens
contaminated with sodium e.g. saline–induced sputum and bronchial washings, are
not suitable for Legionella culture.
Most healthy adults have an antibody titre of < 1:128. Rising antibodies in paired sera
provide good evidence of active infection. Paired sera should ideally be collected two
weeks apart.
Leptospirosis
Specimen:
Serum – Gel
Reference Range: Supplied with report
Since vaccination of cattle was introduced several years ago, the incidence of
leptospirosis has dropped to the point where it is now rare. Host animals include
cattle, pigs, horses and rodents. Human infection typically follows exposure to blood of
infected animals or water contaminated with their urine.
For diagnosis, an antibody screen test is performed followed by (on sera that are
positive) specific agglutination testing against the common strains of leptospira.
After infection, titres rise sharply into the thousands in those who have been infected
in the past. The highest titre may not indicate the cause of the latest illness.
Consultation with the pathologist is suggested when acute leptospirosis is a possibility
so that blood culture in special medium can be arranged.
Leuco–erythroblastic Blood Picture
Immature myeloid cells (myelocytes, metamyelocytes, band neutrophils), and
immature red cells (nucleated red cells), are present due to invasion or disturbance
of the normal marrow.
Causes include:
•
•
•
•
•
•
•
Metastatic malignancy
Primary haematological malignancy
Myelofibrosis
Acute haemolysis
Thalassaemia major, especially after splenectomy
Chronic lymphocytic leukaemia
Chronic myeloid leukaemia.
L
A
Light Chains in Urine
See
Bence Jones Protein (BJP)
Lignocaine
Specimen:Plasma – Lithium heparin
Blood should be collected at least every 12 hours in patients
with evidence of cardiac or hepatic insufficiency.
Reference Range: Supplied with report
Lipaemia
After a fat–containing meal, and in the primary or secondary hypertriglyceridaemias,
the serum is turbid or creamy due to fat particles. A severe degree of lipaemia can
cause technical interference with some tests to the point where they become invalid.
SeeTriglyceride
Lipase
Specimen:
Serum – Gel
Reference Range: < 61 U/L
Increase in lipase levels indicates pancreatitis, and this rise may be seen before amylase.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Lipid Disorders
A lipid screen consists of Cholesterol and Triglycerides. If HDL is required, please
order specifically.
When classifying a lipid disorder for the purposes of treatment, two questions need to
be asked:
1. Is it a primary lipid disorder or secondary to some other metabolic abnormality?
2. Which lipid fractions are elevated?
Cholesterol only
Cholesterol and triglyceride
Triglyceride only.
Secondary disorders
These are easily identified and when treated may entirely correct the lipid
abnormalities.
The common causes are:
•
•
•
•
•
•
•
Obesity
Alcohol
Diabetes
Hypothyroidism
Nephrotic syndrome
Liver disease
Drugs: oestrogens, oral contraceptives, beta blockers, corticosteroids, thiazides,
isotretinoin, antivirals, valproate.
Primary disorders
If no secondary causes are identified the disorder is presumed to be primary. There is
a range of familial disorders which are presumptively identified by the family history or
by physical signs such as xanthomata, corneal arcus.
The commonest familial disorders are familial hypercholesterolaemia and familial
combined hyperlipidaemia. There are many others.
However, the commonest primary dyslipidaemia by a wide margin is polygenic
hypercholesterolaemia, which is not familial. It is identified mainly by exclusion –
no significant family history, no abnormal physical signs. Presumed to result from a
combination of genetic and environmental factors, it provides a convenient diagnostic
refuge.
Which lipid fractions are elevated?
Defining the abnormal fractions is important in both diagnosis and treatment.
The feature of familial hypercholesterolaemia is very high serum cholesterol.
The lipid elevation of familial combined hyperlipidaemia may be cholesterol only (1/3),
triglyceride only (1/3), or both (1/3).
Causes of secondary dyslipidaemias do not reliably produce a constant picture.
For example, though hypothyroidism typically causes elevation of cholesterol only,
it can at times raise the triglyceride also.
L
A
Treatment
Dietary modification is common to the treatment of all dyslipidaemias.
Drug treatment, if required, will depend on whether the abnormality is cholesterol only
(statins being the agents of choice), triglyceride only (fibrates usually first choice) or
both.
PBS Qualifying Criteria for Lipid Lowering Therapy
Patient Category
Lipid Level for PBS Subsidy
Patients with existing coronary disease
Cholesterol > 4mmol/L
Other patients at high risk with one or more of the
following:
• Diabetes mellitus
Cholesterol > 6.5 mmol/L
• Familial hypercholesterlaemia
or
•Family history of coronary heart disease
(first degree relative less than 60 years of age)
Cholesterol > 5.5 mmol/L and
HDL< 1mmol/L
• Hypertension
• Peripheral vascular disease
Patients with HDL < 1 mmol/L
Cholesterol > 6.5 mmol/L
Patients not eligible under the above:
Cholesterol > 7.5 mmol/L
• Men 35–75 years
or
• Post menopausal women up to 75 years
Triglyceride > 4 mmol/L
Cholesterol > 9 mmol/ L
Other patients not included in the above
or
Triglyceride > 8 mmol/L
Lipoproteins
Specimen:
Serum – Gel
In serum, lipids are carried as lipoprotein particles which are grouped in four fractions:
Lipoprotein Fraction
Cholesterol
Triglyceride
Chylomicrons
small
++++
LDL
++++
small
VLDL/IDL
++
++++
HDL
++
small
Capital Pathology Handbook – Interpretation of Laboratory Tests
Diagnosis of the hyperlipoproteinaemias requires a fasting specimen to eliminate the
contribution of dietary triglyceride in both chylomicrons and the VLDL fraction.
Hyperlipoproteinaemias can be familial or secondary (see separate entries under
Triglyceride and Cholesterol for aetiology of the secondary disorders) or, commonly,
a mixture of both. Occasionally the diagnosis is obvious, e.g. the clear–cut familial
hypercholesterolaemias causing severe coronary artery disease in early adult life –
but most of the common hypercholesterolaemias and hypertriglyceridaemias are
part of a large amorphous population, deeply affected by diet and lifestyle and mixed
inextricably with the “normal” population, whoever they are.
Typing of hyperlipoproteinaemias
The WHO classification is that of Fredericksen who recognised six types; I, IIa, IIb, III,
IV, V. The classification has limitations. Familial hyperlipoproteinaemias can present
as different types in different family members. A single aetiology (e.g. hypothyroidism)
can be associated with several types. Treatment can convert one type to another.
Nevertheless the terms are still used. Types II and IV, and to a lesser extent V, are
commonest.
IIa
llb
IV
V
Total cholesterol
+++
+++
+
+
Triglyceride
n
++
+++
+++
LDL cholesterol
++
++
n
n
VLDL cholesterol
n
+
+++
++
Chylomicrons
–
–
++
Turbid serum
–
++
++
++
Note that the classification does not use HDL cholesterol.
SeeApolipoproteins
Lipoprotein Electrophoresis
Cholesterol
Triglyceride
Lipid Disorders
Lipoprotein (a)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Lipoprotein (a), not to be confused with apolipoprotein A which is the protein
component of HDL, is an independent risk factor for coronary artery disease (CAD).
It can be of particular interest in patients whose CAD is not explained by other risk
factors.
Levels are largely genetically determined but tend to rise after the menopause or with
renal impairment. Fasting, exercise or statins have little effect. Levels may be lowered
by niacin and perhaps by fibrates. Acute illness can have an effect and lipoprotein (a)
should not be measured within 3 months of a myocardial infarction.
L
A
Lipoprotein Electrophoresis
Specimen:
Serum – Gel
Four fractions are identified:
Alpha–I
Contains HDL
Pre–beta
Contains VLDL, elevated in Type IV, Ilb or III
Beta
Contains LDL, elevated in Type IIa or Ilb
Chylomicrons
Found in non–fasting serum or Type V
Lipid EPG is a specialist investigation. For most clinical purposes a lipid abnormality is
defined by quantitation of cholesterol fractions and triglyceride on a fasting specimen.
Listeria monocytogenes
Specimen:Blood cultures, CSF, for acute infection
Stool for carrier state.
Listeria, an aerobic gram–positive bacillus, is widely distributed in soil, water and many
animals. It is occasionally found in the human gastrointestinal tract. Spread to humans
is mainly via contaminated food. Food borne outbreaks due to contaminated coleslaw,
milk, soft cheese and mussels have been reported. Although most infections are
mild and harmless, septicaemic infection in pregnant women, presenting as a flu–like
illness, may lead to serious fetal infection leading to stillbirth or neonatal meningitis.
Immunocompromised persons and the elderly are also at risk of invasive disease.
Treatment is penicillin (or amoxycillin) with or without gentamicin. Erythromycin or
cotrimoxazole are less desirable alternatives. Treatment should be for a least 14 days.
L. monocytogenes is resistant to all cephalosporins.
Lithium
Specimen:
Serum – Gel
Desirable therapeutic range: 0.5–1.0 mmol/L
The specimen should be taken 12 hours after the last dose. Steady state levels
are achieved 2–5 days after a dose change, or longer in older patients with renal
impairment. When lithium is stopped, the level falls with a half–life of 1–3 days.
Because of the high incidence of toxic effects, monitoring is regarded as essential
every 3 months, and more often if clinically indicated. Baseline TSH, T4 and creatinine
levels should be measured before commencing therapy and 6 to 12 monthly
thereafter.
Manifestations of toxicity
• A wide range of CNS and GI effects
• Hypothyroidism – relatively common
• Nephrogenic diabetes insipidus
• Serum potassium maybe elevated
• Renal damage with raised s. creatinine may occur when serum levels have been
consistently in the toxic range over a long period
• High serum calcium
• Cardiac toxicity: AV block, T wave changes, premature ventricular contractions.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Drugs that elevate lithium:
• Diuretics
• NSAIDs
• Metronidazole
• ACE inhibitors.
Liver Function Test / Interpretation
Elevated GGT
L
A
Elevated ALP
Predominant Cholestatic Pathology
Capital Pathology Handbook – Interpretation of Laboratory Tests
Predominant Hepatocellular Pathology
Lorazepam (Ativan)
Specimen:
Plasma – Lithium heparin
Reference Range: Supplied with report
Lupus Anticoagulant / Inhibitor
Specimen:
Plasma – Sodium Citrate x3
Reference Range: Not detected
The lupus anticoagulant is an acquired coagulation inhibitor, which is associated
with SLE and thromboembolic disorders. It is associated, but not synonymous with
cardiolipin antibodies. In the context of thrombotic disease or recurrent abortions,
a positive lupus anticoagulant detected on two occasions at least three months
apart, with or without anticardiolipin antibodies, defines an antiphospholipid antibody
syndrome. Anticardiolipin antibodies are sometimes found transiently in healthy
people.
The APTT is prolonged in patients with LAC and fails to correct in the APTT 1+1 test.
The tests performed routinely as the LAC screen are KCT (Kaolin Clotting time),
APTT, PR, DRVVT (Dilute Russell’s Viper Venom Time) and DTTA (Dilute Tissue
Thromboplastin Assay).
L
A
Luteinising Hormone (LH)
Specimen
Serum – Gel
Refernce range:Adult Female
basal mid-cycle post menopausal 2–14 IU/L
14–110 IU/L
15–97 IU/L
Lymphocyte markers
Specimen
Whole Blood EDTA x 2
Lymphocytes play a central role in the immune response. The two main types of cell,
identified by cell marker studies are:
•
•
T cells, primarily responsible for cell–mediated immunity.
B cells, primarily responsible for humoral immunity (immunoglobulin production).
Lymphocytes are also classified by their CD (cluster designation) number.
For example, CD4 cells are T cells used as a marker for immune system damage
in HIV infections.
Lymphocytosis
An absolute lymphocytosis (> 4.0x109/L) is typically found in two situations:
•
•
iral infections, particularly infectious mononucleosis, acute viral hepatitis and
V
CMV infections.
CLL (Chronic Lymphatic Leukaemia), in which the proliferating lymphocytes are
monoclonal B cells with light chain restriction. CLL should always be suspected
when there is an unexplained progressive lymphocytosis at or beyond middle age.
Lymphopenia
Causes include:
•
•
•
•
•
•
Viral illnesses
Pancytopenia due to any cause
Advanced Hodgkin’s disease
Congestive heart failure
Steroid therapy
AIDS.
Variant lymphocytes
These cells, also called reactive lymphocytes, transformed lymphocytes or atypical
lymphocytes, are small lymphocytes which have undergone antigenic stimulation
(e.g. by a virus) that has transformed them into larger cells readily recognised in a
blood film.
The commonest causes of variant lymphocytes are:
•
•
•
•
Infectious mononucleosis (EBV)
Acute viral hepatitis A, B or C
CMV infections
Toxoplasmosis.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Less common causes are:
•
•
•
Other viral infections
Immune disorders
Chronic non–viral infections.
SeeHistopathology
L
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
M
M
A
Macrocytosis
Macrocytes are red cells with an MCV (Mean Cell Volume) above 98. They can be
divided into three groups.
1. The megaloblastic macrocytic anaemias
These are distinguished by the presence of megaloblasts in the bone marrow as
precursors of the peripheral blood macrocytes. Megaloblastic anaemias are due to
B12 or folate deficiency. The macrocytes are oval in shape rather than round. With
well–established deficiency there are hypersegmented neutrophils, neutropenia and
thrombocytopenia.
2. Macrocytosis due to reticulocytosis
Reticulocytes are somewhat larger than older red cells. They are an important
feature of haemolytic and post–haemorrhagic anaemias.
3.Other
In this large group of normoblastic processes, the macrocytes are round. The list
includes:
•
•
•
•
•
Alcoholism
• Myeloma
Liver disease
• Aplastic anaemia
Hypothyroidism
• Myelodysplastic syndromes
Malignant infiltration
• Chronic obstructive airways disease
Cytotoxic drugs (especially hydroxyurea).
Macroglobulinaemia
IgM paraproteins are present in three conditions:
1.Benign IgM paraproteinaemia
The commonest cause. The blood picture is normal, the patient asymptomatic and
IgM levels are static and usually < 10g/L.
2.Waldenstrom’s macroglobulinaemia
A malignant process in which the IgM band is heavy, the blood picture is abnormal
and the marrow infiltrated with abnormal plasmacytoid lymphocytes.
3.Malignant lymphoma or CLL (Chronic Lymphatic Leukaemia)
These should always be searched for clinically, and on the blood film, when an
IgM paraprotein is found.
See Immunoglobulins, IgA, IgG, IgM
Magnesium (Mg)
Specimen:
Serum – Gel
Reference Range: 0.65–1.05 mmol/L
Decreased by:
•
•
•
•
•
•
•
Excessive loss of fluid and electrolytes, diarrhoea etc
Inadequate intake: inadequate parenteral nutrition, low levels in diet and water,
malabsorption
Drugs: diuretics, aminoglycosides, digoxin, cytotoxics, laxative abuse
Alcoholism
Endocrine: hyperthyroidism, hyperparathyroidism, diabetic ketoacidosis, SIADH
Hypokalaemia
Redistribution of Mg into cells, e.g. alkalosis, acidosis, severe illness.
Increased by:
•
•
•
•
Renal insufficiency
Dehydration
Addison’s disease
Haemolysis.
Severe magnesium deficiency affects neuromuscular function and the cardiac
conduction system. Clinically this can be manifested as tetany, convulsions, cardiac
arrest. Epidemiologically, magnesium deficiency in water supplies has been linked to
cardiac dysrhythmias and myocardial infarction.
Causes of Hypomagnesaemia
• Alcoholism
• Renal loss
–Hypercalcaemia
– Renal tubular acidosis
– Osmotic diuresis
• Drugs
– Loop Diuretics
–Gentamicin
–Cisplatin
• Gastrointestinal disorders
–Vomiting
–Diarrhoea
–Malabsorption
• Endocrine
– Diabetes mellitus (probably increased renal loss)
–Hyperaldosteronism
–Hypoparathyroidism
–SIADH
• Miscellaneous
– Acute pancreatitis (sequestration)
– Insulin administration (redistribution)
–Hungry bone syndrome:
post–parathyroidectomy
post–thyroidectomy.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Note
If hypomagnesaemia is associated with a urinary excretion rate > 0.5 mmol/day then
renal wastage is indicated. Levels < 0.5 mmol/day suggest an extrarenal cause.
Developed by N. Walmsley 1995
Malabsorption
The aetiology of malabsorption can be considered under four headings:
• Mucosal defects
– Coeliac disease, the commonest cause
– Crohn’s disease
– Severe lactose intolerance
• Pancreatic disease
– Cystic fibrosis
– Chronic pancreatitis
• Post–infectious malabsorption – Giardia
– Tropical diarrhoea
• Previous gastrointestinal surgery on stomach or small bowel.
Malabsorption particularly involves iron, folic acid, vitamin B12, vitamin K, vitamin D
and fats.
Basic screen tests
• Blood count, looking for macro–or microcytosis
• Iron studies
• B12 and folate
• Stool for Giardia and other pathogens
• Coagulation screen
• Gliadin and TTG antibodies for coeliac disease.
Follow–up tests
• Jejunal biopsy is definitive in the diagnosis of coeliac disease.
M
A
Malaria
Specimen:
Whole blood – EDTA
Malaria is endemic in:
•
•
•
he Pacific west of Fiji. This includes Papua New Guinea, the Solomon Islands,
T
Indonesia. It does not include Fiji, Samoa, Tonga, New Caledonia, Niue, Tahiti or
Hawaii.
SE Asia and India
Africa and S. America.
The diagnosis needs to be considered in any visitor to these areas with unexplained
fever, sweats, headache, malaise, anaemia, abnormal liver enzymes.
Diagnosis is established by detecting parasites in thick and thin films. Serial blood
specimens may be required. Leucopenia is usual but there can be leucocytosis.
Variant lymphocytes are occasionally seen. Malaria can be accompanied by anaemia
which may be caused by a haemolysis, marrow depression or hypersplenism.
P. falciparum causes severe, sometimes lethal, illness.
P. vivax is less severe but can relapse.
P. ovale and P. malariae are the two less commonly found subtypes.
Treatment should be discussed with those who have experience in treating malaria.
Malassezia furfur
A fungus causing tinea versicolor (pityriasis versicolor) which show as brown scaly
patches on white skin or pale patches on dark skin. Microscopy of skin scrapings
shows grape–like clusters of cells. The organism is not readily cultivated. Diagnosis is
based on clinical features, direct microscopy and absence of fungal growth on culture.
Treatment is with a topical imidazole, ciclopiroxolamine or terbinafine. Oral terbinafine
is not effective.
Mantoux Test
Method
The standard Mantoux test consists of an intradermal injection of 10 units of tuberculin
(PPD–purified protein derivative) contained in 0.1 mL of solution. The injection is made
into the anterior aspect of the forearm and read three days later (2–4 days are in the
outer limits if 3 days is impracticable).
When reading the test, the arm is palpated to define an area of induration and the
diameter of this induration, if present, is measured in mm transverse to the long axis of
the forearm.
An area of surround erythema (redness), or erythema alone, is ignored.
If vesiculation (blistering) or necrosis (darkening or ulceration) of the indurated skin is
present, these must be described as an important part of the result.
Interpreting results
A diameter of induration > 10 mm is described as positive and is consistent with
current or past TB infection.
Capital Pathology Handbook – Interpretation of Laboratory Tests
A strong positive in a person with active TB disease may be > 15 mm with vesiculation
and necrosis.
Previous BCG vaccination is associated with induration up to 15 mm though
usually < 5 mm.
In immunocompromised persons, the tuberculin reaction may be reduced or
suppressed, even in the presence of proven infection.
False positive Mantoux results
These are usually due to operator error, particularly reading errors where the area of
erythema rather than induration has been read.
False negative Mantoux results
Although a negative Mantoux usually indicates that a person has never been exposed
to TB, there are important exceptions:
•
•
•
•
•
The person had TB in the past but the immune response has faded
Overwhelming infection or illness of any type, including overwhelming TB
Patients with immune suppression – HIV, immunosuppressive drugs, sarcoidosis,
renal failure, malignancy, malnutrition
Acute viral infections, recent live virus vaccinations
Neonates.
Measles Antibodies (morbilli, English measles, rubeola)
Specimen:
Serum – Gel
Reference Range: Supplied with report
The test can be for immune status (IgG antibody) or acute illness (IgM).
IgM antibodies appear about two days after the rash and peak about 2 weeks later.
IgM antibodies disappear after the acute illnesses. IgG antibodies persist for life.
Meloidosis Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
MEN (Multiple Endocrine Neoplasia)
These are familial clusters of endocrine adenomas or hyperplasias:
•
Type IParathyroid adenoma (95%)
Pancreatic islet cell adenoma or hyperplasia (80%)
Pituitary adenoma (50%)
•
Type IIMedullary thyroid carcinoma (95%)
Phaeochromocytoma (50%)
Parathyroid adenoma/hyperplasia (40%).
Meningitis
Please see Neisseria meningitidis.
M
A
Mercury
Specimen:24–hour urine (nil preservative) or random urine
Whole blood – Trace element tube.
Reference Range: Supplied with report
Most of the public interest in mercury toxicity centres on amalgam in teeth but
studies have not shown harmful effects. After an exhaustive investigation, the U.S.
Public Health Services concluded there is no serious health risk. Urine mercury
levels in people with filled teeth are less than 5% of the stated acceptable limit.
Organic mercury compounds have been used as fungicides with widespread
poisoning in Iraq in 1971 when bread was accidentally made from seed wheat
preserved with methyl mercury. Mercury–contaminated waste in sea–water is passed
up the food chain to reach its most concentrated levels in large predatory fish such
as shark, tuna and swordfish. The Miranda Bay disaster in Japan in 1955 was due to
industrial waste discharge being concentrated in edible fish.
Mercury as calomel in children’s teething powders and laxatives was found to be the
cause of acrodynia (pink disease) as late as the 1940s.
Mercury poisoning causes renal damage and neurotoxicity.
Methadone Screening
Specimen:
Random urine (nil preservative)
Reference Range: Not detected
See
Drug Screen
Methyltetrahydrofolate Reductase Gene (MTFHR gene)
Specimen:
Whole blood – EDTA
SeeThrombophilia
Microalbumin
SeeProteins, urine
Albumin Excretion Rate (AER)
Microsomal Antibodies
Specimen:
Serum – Gel
Reference Range: Not detected
SeeThyroid Antibodies
Capital Pathology Handbook – Interpretation of Laboratory Tests
Molecular Genetics – Genetic Studies
Increasing numbers of inherited diseases are now identifiable using genetic studies.
Tests are available or are being developed for:
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Fragile X syndrome
Myotonic dystrophy
Prader Willi syndrome and Angelman syndrome
Huntington disease
Spinocerebellar ataxia
Familial adenomatous polyposis
Multiple Endocrine Neoplasia Type 2
Spinocerebellar muscular atrophy
Spinal muscular atrophy
Duchenne muscular dystrophy
Cystic fibrosis
Breast cancer testing
Adrenoleukodystrophy
Hereditary haemochromatosis.
Morphine Screening (drug screen)
Specimen:
Random urine (nil preservative)
Reference Range: Not detected
See
Drug Screen
Mumps Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Interpretations:
IgM
IgG
Current or recent infection
+
+
Previously infected, now immune
–
+
Never infected
–
–
The diagnosis of mumps is usually based on clinical grounds and does not necessitate
a blood test.
Serum amylase levels rise sharply.
M
A
Myasthenia Gravis
Specimen:
Serum – Gel
This is an autoimmune disorder in which neuromuscular transmission is reduced to
a varying extent by acetylcholine receptor autoantibodies (anti–ACLR) which are
detectable in about 80% of cases.
Diagnostically the Tensilon (edrophonium) test is used, an injection of
anticholinesterase which restores neuromuscular function.
See
Acetylcholine Receptor Antibody
Mycobacterium avium (MAC)
Specimen:Sputum
Mycobacterium avium complex (MAC) infection is seen predominantly, but not
exclusively, in patients with HIV infection. In that group it usually presents with
dramatic fever spikes, rigors, malaise, often hepatosplenomegaly, anaemia and
cholestatic liver function tests.
Otherwise normal children may also present occasionally with cervical adenopathy
due to MAC. It is sometimes recovered from sputum specimens sent to TB culture.
Although it can cause true respiratory tract disease it is often just a coloniser in an
abnormal respiratory tract. Evidence for its being a pathogen includes repeated
isolation, a positive smear result and CXR changes. Treatment regimens include
ethambutol and clarithromycin with or without rifabutin. Decisions on whether to treat
and with what, require specialist advice.
Mycoplasma pneumoniae
Specimen:
Paired sera – Gel
Mycoplasma is a bacterium not demonstrable with usual stains or growth media. It
is the commonest cause of atypical pneumonia, particularly in children and young
adults, and can spread as a low–grade epidemic. It responds well to erythromycin or
tetracycline but not to penicillin.
Diagnosis is by exclusion, by response to therapy, and by demonstrating a rising
titre in paired sera, the first collected as soon as possible in the illness, the second 2
weeks later. Not infrequently the antibody titre has already plateaued by the time the
first specimen is collected.
SeePneumonia
Myelodysplastic Syndromes (MDS)
Also known as evolving leukaemia or preleukaemia.
These conditions are primary bone marrow disorders which usually behave as slowly
evolving bone marrow failure syndrome. Most often they present insidiously and in
an older population. A variety of peripheral blood findings may be noted including
pancytopenia, refractory anaemia, macrocytosis with anaemia, neutropenia and
thrombocytopenia. Diagnosis is by excluding vitamin deficiency (B12, folate) and
demonstrating dysplastic features in the bone marrow. Chromosome analysis and cell
Capital Pathology Handbook – Interpretation of Laboratory Tests
marker studies may also be performed on the bone marrow. Management is usually
by supportive care but in selected patients more aggressive therapy may be indicated.
Myelofibrosis
Myelofibrosis is a chronic myeloproliferative disorder characterised by bone marrow
fibrosis, extramedullary haemopoiesis, splenomegaly and a leucoerythroblastic
blood film. The condition may present as myelofibrosis at the outset or be the end
stage of another myeloproliferative disorder such as polycythaemia vera or essential
thrombocytopenia. Survival varies with a median of four years. Therapy includes blood
transfusions, cytoreductive therapy (usually hydroxyurea) for thrombocytosis, and
splenectomy in selected cases.
See Myeloproliferative Disorders
Myeloproliferative Disorders (MPD)
These conditions are characterised by excessive production of erythroid, myeloid
and megakaryocytic cell lines (panmyelosis). In many situations only one cell line
may predominate, e.g. platelet proliferation causing essential thrombocythaemia. In
polycythaemia rubra vera (primary polycythaemia), erythroid hyperplasia is the most
prominent feature with variable increases in myeloid cells (neutrophils) and platelets.
There may also be excessive proliferation of reticulin and fibroblasts within the bone
marrow giving rise to myelofibrosis. Acute myeloid leukaemia can develop as a
terminal event, usually untreatable, in any of the MPDs.
Myoglobin
Specimen:
10 mL random urine
Reference Range: Not detected
Myoglobinuria occurs after skeletal muscle trauma or myocardial infarction (MI). After
major trauma, myoglobin imparts a coffee colour to urine but small amounts, as in MI,
are detectable only by chemical tests.
SeeTroponin
Mysoline (Primidone)
Specimen:
Plasma – Lithium heparin
Reference Range: Supplied with report
Mysoline is metabolised to phenobarbitone which is responsible for much of the drug’s
anticonvulsant activity. Mysoline drug levels are monitored as phenobarbitone.
M
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
N
N
A
Needlestick Injuries
Investigations
Bloods from the exposed workers and the source specimen should be tested for
markers of infection and immunity.
•
•
•
•
HIV antibody/antigen
HBs antigen
HBs antibody
HCV antibody.
The event must be fully documented and the exposed worker counselled and kept informed.
Prevention
All health care workers should be immunised against HBV.
Venepuncturists and nurses giving injections should be trained in safe practice.
Needles and other blood–contaminated sharps must be disposed of into punctureproof containers.
Neisseria
These gram–negative diplococci are typically found inside neutrophils in direct smears
from lesions. The two main pathogens are N. gonorrhoeae and N. meningitidis.
N. catarrhalis, a member of the normal flora of the throat, was known as Branhamella
catarrhalis and is now known as Moraxella catarrhalis. It is a recognised respiratory
tract pathogen.
A. Neisseria gonorrhoeae
Specimen:Bacterial swabs must be placed in transport medium immediately
after collection. Store room temperature, not in the fridge, and
transport to the laboratory as soon as possible–they will survive
24 hours in transport medium. In general, Neisseria are sensitive
to drying, sunlight, heat, cold and many disinfectants. PCR
is available and can be done off fist void urine and ThinPrep.
Please indicate specific testing required on request form.
The main infections are:
Male STDUrethritis and sometimes epididymitis. The organism can
also sometimes be recovered from the rectum and throat.
Female STDEndocervicitis is the initial infection with variable spread
upwards to fallopian tubes and downwards to urethra and
perianal skin. As with males, gonococci also grow in the
throat and rectum.
Neonatal conjunctivitisAcquired during birth from an infected mother.
Gonococcal arthritisA suppurative arthritis. Synovial fluid aspirate establishes
the diagnosis.
Treatment
As with any STD, treatment of partners is essential. Concomitant infection with
Chlamydia is common and empirical treatment for this is recommended e.g. Ig
azithromycin stat.
B. Neisseria meningitidis
The causative organism of meningococcal meningitis and septicaemia which can
occur sporadically or in epidemics. Travellers to countries where epidemics occur,
e.g. Nepal, should be vaccinated according to recommendations current at the time of
travel. There are three main sub–groups: A, B and C.
Suspected meningococcal meningitis is a medical emergency and in the community
situation, parenteral antibiotics should be given by the diagnosing doctor before
sending the patient to hospital.
Neutrophil Alkaline Phosphatase (NAP score)
Specimen:
Whole blood – Lithium Heparin
Reference Range: Supplied with report
Films must be made within a few hours of collecting the specimen. Although the test is
of low specificity it is of some value in the following conditions:
Elevated NAP
Reduced NAP
• Polycythaemia vera (NAP up to 300)
• Myelofibrosis
• Infections
• Other inflammatory conditions
• Pregnancy.
• Chronic myeloid leukaemia
• PNH.
Neutrophil alkaline phosphatase activity is present in the specific granules of all
myeloid cells with highest activity in the youngest neutrophils.
Neutrophils (Polymorphs)
Neutrophil counts are expressed and interpreted in terms of the absolute count rather
than % of total white cell count.
Neutropenia
The adult reference range is 2.0– 8.0x109/L.
Mild neutropenia, 1.0– 2.0x109/L, is a common finding, often suspected to be caused by
an immune–medicated mechanism (immune neutropenia) when drugs are not implicated.
Below 0.5x109/L there is an increase in spontaneous infection with a dramatically
increased risk below 0.2x109/L (=agranulocytosis).
Common causes of neutropenia
• Viral infection, sometimes with an associated thrombocytopenia
• Some bacterial infections, e.g. typhoid
• Overwhelming sepsis, often with toxic granulation of neutrophils and a shift to the left
• B12 or folate deficiency
• Aplastic anaemias
• Malignancy – haematological malignancy or metastatic deposits in bone marrow
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
•
•
•
Immune – mediated – post viral, Felty’s syndrome, SLE
Hypertension
Thyrotoxicosis
Chronic benign neutropenia, including cyclical neutropenia.
Drugs those more commonly implicated are:
•
•
•
•
•
•
•
•
Phenylbutazone, indomethacin
Chlorpromazine, promazine, other phenothiazines
Carbamazepine
Propylthiouracil, carbimazole
Sulphonamides, cotrimoxazole
Dapsone
Thiazides
Captopril.
Cyclical neutropenia
Characterised by periodic fluctuations in the neutrophil count with the nadir occurring
usually at three weekly intervals. In the most severe cases the neutropenia may be
extremely low, < 0.2x109/L, with infective complications, malaise and fever. Hospital
admission may be required, as occasionally these may be life–threatening events.
In some patients, due to the severity of the neutropenic episodes and the infective
complications, growth factor support with granulocyte colony stimulating factor (G–
CSF) is required.
Neutrophilia
A neutrophil count > 8. 0x109/L (in non-pregnant adults)
Minor degrees of neutrophilia are a common response to almost any disease process.
In more serious disorders, neutrophil precursors may be present, the sequence
from cell division to maturity being myeloblast, myelocyte, metamyelocyte, band
neutrophil, segmented neutrophil. An increase in band neutrophils, meta–myelocytes
and eventually myelocytes, i.e. a “shift to the left”, should prompt a search for serious
disease.
Causes of neutrophilia
• Bacterial infection – there may be a shift to the left and with more severe infections
there may be toxic granulation and eventually neutropenia as the marrow is
overwhelmed. At the other end of the scale is the leukaemoid reaction, a massive
outpouring of cells in response to infection with counts up to 80x109/L and a
striking shift to the left.
• Tissue injury – due to infarction, burns, surgery, trauma and other processes
causing necrosis
• Other inflammatory disorders – connective tissue disorders, etc
• Malignancy
• Drugs – steroids, epinephrine, heparin
• Other – haemorrhage, stress, metabolic disorders.
N
A
Nipple Secretions for Cytology
These specimens should be collected by applying the slide directly to the nipple and
smearing secretion onto the slide.
If the secretion is scant the slide can be lightly touched against the nipple several
times.
A mixture of air–dried and wet–fixed slides is appropriate. Slides can be fixed using
a spray fixative or by immersion in 95% alcohol for 20 minutes immediately after
collection. Air–dried slides should be dried rapidly (rapidly waving the slide backwards
and forwards through the air ensures rapid drying). The slides must be marked as to
whether they are air–dried or wet–fixed, as laboratory preparation and staining differ
for each.
Labelling the slides on the frosted end of the slide with pencil, indicating the patient’s
family name, first name and date of birth.
Capital Pathology Handbook – Interpretation of Laboratory Tests
O
O
A
Oestradiol (E2, Oestrogen)
Specimen:
Serum – Gel
Reference Range: Supplied with report
•
•
Oral contraceptives are associated with very low levels
Pregnancy is associated with very high levels.
The assay is specific for oestradiol. Other oestrogens such as ethinyl oestradiol or
conjugated equine oestrogens (Premarin) are not detected.
Oestradiol is the major endogenous oestrogen is ovulating women. It rises to a pre–
ovulatory peak mid–cycle, falls, and then returns to a luteal phase plateau before
falling again prior to menstruation.
Indications for estimating oestradiol are strictly limited:
•
•
•
During artificial induction of ovulation
Investigation of feminising states including precocious puberty, gynaecomastia
Investigation of possible pituitary–gonadal deficiency states in women.
It is not used during the early investigation of infertility, for diagnosing menopause,
for monitoring HRT (hormone replacement therapy), or for investigating irregular
menstruation.
Oestriol (E3)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Unconjugated serum oestriol is used when testing for Down’s syndrome during the
first or second trimester.
See Pre-Natal Testing
Opiates Screening (Drug Screen)
Specimen:
Random urine (nil preservative)
Reference Range: Not detected
See
Drug Screen
Osmolality
Specimen:Serum – Gel
Urine, early morning, or random, or timed collect, or 24–hour urine.
Reference Range:Serum
Urine
275–295 mosmol/kg water
50–1500 mosmol/kg water
Urine osmolality, which is under the control of pituitary ADH, varies over a wide range
to ensure that alterations in fluid intake have little effect on the tightly controlled serum
osmolality. Urine osmolality is typically highest on rising in the morning because of the
absence of fluid intake during sleep. An osmolality > 600 (especially > 800) makes
diabetes insipidus unlikely.
An approximate serum osmolality is given by the formula:
Osmolality = 2 x Na+ + urea + glucose
The normal physiological response to hyponatraemia is the secretion of dilute urine
with osmolality < 100. A higher osmolality suggests SIADH.
Osteocalcin (Serum Bone GLA protein)
Specimen:Serum – Gel
Spin, separate and freeze serum immediately.
Suggest collection of specimen at your nearest Collection Centre.
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
P
P
A
Paget’s Disease of Bone
Asymptomatic Paget’s is a common cause of elevated ALP (150–500 units or higher)
in the elderly and the ALP level is an indication of disease activity and response
to treatment. 24–hour urine N–telopeptide is another indicator used in monitoring
treatment.
A bone scan is a sensitive method for identifying foci of Paget’s disease.
See Alkaline Phosphatase (ALP)
Pancytopenia
A reduction in all three cellular elements in blood i.e. anaemia, leucopenia
(neutropenia) and thrombocytopenia. An unexplained pancytopenia is always an
indication for a bone marrow biopsy.
Causes include:
• Haematological disorders requiring bone marrow examination – aplastic anaemia,
leukaemias, myeloma, myelodysplastic disorders, metastatic tumour
• Drug effects
• B12 or folate deficiency
• Hypersplenism
• SLE, PNH
• Overwhelming infection, HIV.
PAP Smear
The quality of the Pap smear specimen is of critical importance if false negative
cytology reports are to be minimised.
There are two main reasons for the occurrence of false negative Pap smear reports.
One is laboratory error and the other is sampling error. At Capital Pathology we have
put in place numerous quality assurance measures to minimise laboratory false
negatives. Sampling errors can be reduced by taking an optimal sample. This also
helps the laboratory in the interpretation of the smear.
See
Cervical Cytology
Paracetamol (Acetaminophen)
Specimen:Serum – Plain clot or Lithium heparin
Do not use gel tube.
Serum paracetamol levels are used to assess the need for N-acetylcysteine
administration in all patients with deliberate paracetamol self-poisoning, regardless of
the stated dose.The recommendations for the management of paracetamol poisoning
in Australia and New Zealand are derived from consensus guidelines, that include the
treatment nomogram shown.
The nomogram uses a single line to simplify decision making, and it uses the
treatment threshold with the most clinical data to support its efficacy and safety.
New treatment nomogram for paracetamol poisoning
Daly FFS, Fountain JS et al. Guidelines for the management of paracetamol poisoning in Australia and
New Zealand — explanation and elaboration. Med J Aust 2008; 188(5): 296-302. © Copyright 2008.
The Medical Journal of Australia – reproduced with permission.
Parathyroid Hormone (PTH)
Specimen:Serum – Gel
Spin, separate and refrigerate serum.
Freeze if to be kept overnight.
Reference Range: 1.0–6.0 pmol/L
Elevations of PTH with elevated calcium:
•
•
•
rimary hyperparathyroidism due to solitary adenoma (85%) or hyperplasia (15%)
P
is the commonest cause.
Lithium therapy
Familial benign hypocalciuric hypercalcaemia is a rare autosomal dominant
disorder in which excessive renal calcium reabsorption is associated with relatively
small elevations in PTH. Parathyroid surgery is contraindicated.
Elevations of PTH without elevated calcium:
•
•
Renal failure–2° hyperparathyroidism
Vitamin D deficiency.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Paroxysmal Nocturnal Haemoglobinuria (PNH)
Specimen :
Whole blood – EDTA x 2
Reference Range: Supplied with report
An absence of CD59 activity on the cell surface of red cells characterises Paroxysmal
Nocturnal Haemoglobinuria (PNH).
PNH is a rare, acquired haematological disorder usually presenting in adult life.
Its features include:
•
•
•
Intravascular haemolysis
Venous thrombosis
Marrow hypoplasia.
PNH is due to an acquired mutation giving rise to a clone of abnormal stem cells
affecting erythrocytes, leucocytes and platelets.
The haemolysis, though chronic, is intermittent, occasionally paroxysmal with severe
anaemia. It is mediated by complement and is made worse by reduction in blood
pH as occurs at night, during exercise, or in vitro – as in Ham’s test which is used in
diagnosis.
The nonspecific laboratory abnormalities include:
•
•
•
Anaemia, neutropenia, thrombocytopenia
Reduced haptoglobins, reticulocytosis
Haemoglobinuria, haemosiderinuria.
More specific tests for PNH are:
•
•
•
•
Absence of CD59 activity on the erythrocyte cell surface
Ham’s acidified serum lysis test
Sucrose lysis test
CD59.
Pasteurella multocida
A gram–negative bacillus that is isolated from skin lesions. The organism is a common
inhabitant in the mouths of cats and dogs and infection is frequently associated with
animal bites or scratches. Regional lymphadenopathy, chills and fever can result. It is
susceptible to penicillin and amoxycillin. Wound debridement may also be necessary.
Paternity Testing
Specimen:Buccal swab
DNA testing can provide parentage information for family reasons, immigration and
forensic purposes.
Please note that these tests are not covered by Medicare.
For further information, please contact the Doctors Service Centre on 02 6285 9803.
P
A
PCR (Polymerase Chain Reaction)
A molecular method to amplify genetic material. The amplified sequence can then be
detected by a variety of methods. Often used to detect small numbers of organisms in
a specimen, e.g. Toxoplasma gondii in amniotic fluid or M. tuberculosis in CSF. PCR
is also used to amplify sequences for human gene studies, e.g. paternity testing and
some inherited genetic disorders.
Pemphigus Antibodies
Specimen:
Serum – Gel
Reference Range: Not detected
Peripheral Neuropathy
Pathologies to consider include:
•
•
•
•
•
•
•
•
•
•
•
•
Diabetes
Viral infection: Guillain–Barré
Alcoholism
Connective tissue disorders
Paraproteinaemias
Toxins: lead
Drugs: amiodarone, phenytoin, nitrofurantoin and others
Nutritional disorders
Vasculitis
Deposition diseases e.g. amyloid
Para-neoplastic
Hereditary.
Pernicious Anaemia
An autoimmune disease, rare before the age of 40, causing destruction of gastric
parietal cells and intrinsic factor and hence reduced absorption of vitamin B12.
Laboratory features include some or all of the following:
•
•
•
•
•
•
•
•
•
•
Anaemia, thrombocytopenia, neutropenia
Serum vitamin B12 reduced
Blood film features – oval (not round) macrocytes
– hypersegmented neutrophils
Macrocytosis – MCV up to 120
Intrinsic factor and/or parietal cell antibodies
– 90% are positive for one or
both of these
Folate – serum levels normal or increased
Bone marrow – megaloblastic change
Prominent increase in reticulocytes 10 days after commencing therapy with B12.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Pesticides
Specimen:Whole blood – Lithium heparin
Urine 20 mL random collection.
Reference Range: Supplied with report
Phenobarbitone
Specimen:Serum – Gel
Trough level is taken just before next dose (within one hour).
Peak level is taken 1–3 hours post dose.
Reference Range: Supplied with report
The half–life of phenobarbitone is 4 days.
SeeAnticonvulsants
Phenylalanine
Specimen:Blood spots on Guthrie card
Plasma – Lithium heparin for new born screening.
Monitoring PKU–quantitative phenylalanine levels is also done on blood spot
specimens.
Phenytoin
Specimen:Serum – Gel
Trough level is taken just before next dose (less than one hour).
Peak level is collected between 4–7 hours post dose.
Reference Range: 40–80 umol/L therapeutic range adults
SeeAnticonvulsants
P
A
Phosphate
Specimen:
Serum – Gel
Reference Range: Adults 0.80–1.50 mmol/L
Phosphate, also called inorganic phosphorus, is used diagnostically in two main
situations:
1.Hypercalcaemia – levels are reduced or low normal in primary
hyperparathyroidism and vitamin D deficiency, but are usually increased or normal
in other hypercalcaemias
2. Renal failure – phosphate is elevated.
A wide range of other disorders cause hyper–or hypophosphataemia but diagnostic
value is limited.
Phosphate levels can be falsely raised if the specimen is haemolysed.
Causes of Hypophosphataemia
•
•
•
•
•
•
•
•
•
Possible causes:
Hypercalcaemia
Associated with hyperparathyroidism or malignancy
Aluminium antacid therapy
Hyperalimentation
Nutritional recovery
Alkalaemia
Treated diabetic acidosis
Alcoholism
Hypomagnesaemia.
Causes of Hyperphosphataemia
•
•
•
•
•
•
•
•
Possible causes:
Artefactual
Haemolysis
Delayed separation of plasma from RBC
Renal failure
Phosphate rarely exceeds 4 mmol/L
Consider additional causes if phosphate > 4 mmol/L
Infancy and childhood (Higher reference ranges)
Increased intake
Vitamin D excess
IV therapy
Cell leakage
Acidosis
Hypoparathyroidism.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Platelet Antibodies
Specimen:
Serum – Gel and Whole blood – EDTA
Reference Range: Supplied with report
Platelet antibody tests are most often performed in the investigation of an isolated
thrombocytopenia. Both platelet–associated (PAA) and serum platelet (SPA)
antibodies are measured. A positive result for either test supports an immune aetiology
though immune thombocytopenia may be present in patients with negative PAA and
SPA.
Platelet Count
Specimen:
Whole blood – EDTA
Reference Range: 150–400 x 109/L
Reduced platelets – see Thrombocytopenia
Raised platelets –
see Thrombocytosis
Platelet Dysfunction
This is suspected in patients presenting with a bleeding history and a normal platelet
count, normal coagulation factor screen but prolonged bleeding time. The term
“minimal bleeding disorders” is sometimes used. Occasionally, platelet aggregation
and other studies are used where further investigation is required.
Platelet function disorders can be:
Congenitalrare defects of aggregation, adhesion or the platelet
release reaction.
• Acquired – drug effects, e.g. NSAIDs
– haematological malignancies
– immune thrombocytopenia (ITP)
– myeloproliferative disorders
– renal failure.
•
Pneumocystis carinii
Pneumocystis carinii was considered to be a protozoan parasite but genetic studies
suggest it is most likely related to the fungi.
It causes an acute to sub–acute, often fatal, pulmonary disease in the
immunocompromised. It is a major disease problem in those infected with HIV.
Diagnosis is by detecting organisms in bronchial brushings, open lung biopsy and lung
aspirates. A variety of stains may be used to show the organism.
Treatment is high dose cotrimoxazole and should be discussed with those who have
experience in treating this condition.
P
A
Pneumonia
At the time of clinical diagnosis an attempt, often unsuccessful, should be made
to obtain a sputum sample. Culture may show Strep. pneumoniae in classical
lobar pneumonia, or Haemophilus influenzae and/or S. pneumoniae in pneumonia
associated with chronic bronchitis.
Antibiotic therapy is usually started immediately on an empirical basis. A sputum
specimen obtained after antibiotic treatment has started, will not be useful.
Atypical pneumonias
These are almost as common as the classical bacterial pneumonias.
•
•
•
•
•
•
ycoplasma pneumoniae is the commonest particularly in children and
M
young adults
Psittacosis affects those who have occupational contacts with birds,
including poultry
Legionnaire’s disease – See Legionella antibodies
Q fever
Brucella
Influenza.
For initial investigation (screen), the following tests are recommended:
serology for Mycoplasma pneumoniae, Q fever, Legionella, Chlamydia pneumoniae,
Chlamydia psittaci and influenza.
SeeChlamydia psittaci
Chlamydia pneumoniae
Legionella antibodies
Mycoplasma pneumoniae
Q Fever antibodies
Polycystic Ovary Syndrome (PCOS)
This common endocrine disorder is characterised variably by:
•
•
•
•
•
•
•
•
•
•
•
Amenorrhoea or oligomenorrhoea
Infertility / anovulation with oestrogen present
Hirsutism, acne, alopecia
Mildly elevated testosterone – produced by the ovaries
Increased extra–ovarian production of oestrogen
LH elevated, FSH depressed, LH/FSH ratio > 2
Prolactin elevated
Obesity
Ovaries enlarged with multiple cysts visible on ultrasound
Insulin elevated due to insulin resistance
Family history of PCOS.
Not all these features are necessarily present. The combination of androgen excess
and obesity leads to increased extra–ovarian oestrogen production which increases
LH and decreases FSH.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Polycythaemia
Defined as a haemoglobin concentration above the reference range for age and sex.
The approach to diagnosis is to distinguish between polycythaemias due to an
increased red cell mass (primary and secondary polycythaemias) and those due to a
reduced plasma volume (relative polycythaemias).
1. Polycythaemia rubra vera (primary erythrocytosis)
A myeloproliferative disorder usually occurring in middle or older age in which red
cells, and often white cells and platelets, are increased.
Laboratory features
• Red cells – Hb and PCV are increased.
• The red cell mass is elevated (usually > 36ml/kg in males) but the plasma volume
is normal. By convention the packed cell volume is used to monitor the treatment
response.
• White cells – there is an increase in neutrophils often with a shift to the left with
band neutrophils, metamyelocytes and occasionally myelocytes.
• Platelets – 400–800x109/L in most cases with counts over 1000x109/L
occasionally. Giant forms may be present.
• NAP score – increased in 70%.
2. Secondary polycythaemia
Increase in the red cell mass due to tissue hypoxia caused by:
•
•
•
Chronic respiratory disease
Congenital heart disease
High altitude.
Secondary polycythaemia also occurs in some renal disorders, particularly tumours,
where there is increased erythropoietin production.
3. Relative or “stress” polycythaemia
The commonest form of polycythaemia, the elevated haemoglobin being secondary to
a depletion in the plasma volume. By definition the red cell mass (volume) is normal.
Clinical associations are:
•
•
•
•
•
Smoking
Alcohol
Stress
Dehydration
Diuretics.
This condition is diagnosed by demonstrating a normal red cell mass and a reduced
plasma volume.
P
A
Polydipsia / Polyuria
Urinary frequency due to UTI (urinary tract infection) or lower urinary tract problems,
can be distinguished from polyuria by collection of a 24 – hour specimen to document
urine volume.
Causes of polyuria include:
•
•
•
•
•
•
iabetes mellitus (random glucose levels usually above 15 when polydipsia/
D
polyuria are present)
Hypercalcaemia
Hypokalaemia
Lithium therapy
Psychogenic polydipsia (compulsive water drinking)–not uncommon
Diabetes insipidus.
To differentiate between the latter two, see Diabetes insipidus.
Polymyalgia rheumatica
A relatively common disorder, particularly in the elderly, characterised by distressing
shoulder and pelvic girdle pain originating from muscles rather than joints. It is often
associated with temporal arteritis.
ESR and CRP are almost invariably (but not always) raised.
Response to steroids is dramatic.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Porphyrias
Specimens:All specimens must be protected from light and delivered to the
lab as soon as possible.
Urine:During an attack of suspected acute porphyria, collect a random
urine for PBG
24–hour urine.
Faeces:
Casual specimen.
Red cells:
EDTA tube
There are two main syndromes:
Acute porphyria
• Abdominal pain (autonomic neuropathy)
• Peripheral neuropathy
• Neuropsychiatric manifestations.
Attacks are provoked by drugs that induce liver enzymes – barbiturates, alcohol, oestrogens,
sulphonamides, anticonvulsants, and others. In women, neuropathic symptoms may be
cyclical, associated with oestrogen peaks.
Cutaneous porphyria
Presents with photosensitivity and blistering in sun–exposed areas, notably backs of
hands, forearms, face, where accumulated porphyrins react with light to produce skin
damage.
Symptoms are provoked and exacerbated by oestrogen, alcohol, and iron supplements.
Investigations
During a suspected acute attack, e.g. recurrent acute abdominal pain not otherwise
explained, a fresh urine sample should be taken to the lab urgently for PBG.
During latent phases of acute porphyrias, abnormalities may be undetectable.
Cutaneous photosensitivity is most often due to PCT (Porphyria Cutanea Tarda) which is
diagnosed by increases in urine and faecal porphyrins. Diagnosis of the rare erythropoietic
porphyrias requires a blood specimen.
P
A
Potassium (K+), serum
Specimen:
Serum – Clot or gel
Reference Range: Adults 3.5–5.5 mmol/L
Hyperkalaemia
Hypokalaemia
Capital Pathology Handbook – Interpretation of Laboratory Tests
Pre–Natal Testing
1. First trimester screening
A test is available to calculate the risk of Down’s Syndrome and neural tube defects
in the first trimester. The first trimester Down’s Syndrome risk assessment combines
measurement of serum free beta HCG (free BHCG), serum pregnancy associated
placental protein A (PAPP–A), alphafetoprotein and oestridiol and measurement by
ultrasound of nuchal fold thickness.
Together these parameters are able to give an adjusted risk for Down’s Syndrome.
The information from first trimester screening can be presented in a number of ways,
however the maximal detection rate is achieved through the combined assessment of
the biochemical markers and nuchal thickness.
Patient requirements for combined first trimester screen:
a)Serum for biochemistry collected from between 9 weeks until 14 weeks,
2 days inclusive.
b) Ultrasound performed from 11 to 13 weeks gestation.
The final combined risk assessment is provided by the radiology practice who
performed the ultrasound.
2. Second trimester screening
The combination of maternal serum alpha–feto protein (AFP), unconjugated oestriol
(uE3) and human chorionic gonadotrophin (HCG) taken at 15 to 20 weeks gestation
together with gestational age, weight, maternal age, the presence or absence of
diabetes and the previous obstetric history can be used in a calculation to obtain a risk
assessment for fetal Down’s Syndrome and open Neural Tube Defects.
An accurate estimate of gestational age is critical as circulating levels of all three
serum markers vary with gestational age.
Assay results which may appear abnormal (and therefore high risk) for a patient with
a gestational age of 18 weeks may be normal and therefore, low risk if, following
ultrasound, the gestational age is determined to be 15 weeks.
Note: New or additional information can be put into the computer to give another
assessment using the same assay value if, for instance, the gestational age is found
to be different to that originally stated on the request form. No charge is made for this
additional service.
The test is solely a SCREENING TEST and indicates the need for a more specific test
(ultrasound, amniocentesis or chorionic villous sampling) to determine the outcome of
the pregnancy.
The cut–off for a “screen positive” result requiring further assessment is arbitrarily set
at 1 in 250. At a cut–off of 1:250, the test will identify approximately 60% of Down’s
Syndrome babies.
Neural tube defect risk assessment uses the AFP level to calculate the risk.
P
A
Comparison of Antenatal Screening Methods
Screening test
Gestation
(weeks)
Down’s Syndrome
detection rate (5%
FPR)
First trimester serum
Biochemistry (free beta–HCG and
PAPP–A) (FTS)
9–14 / 2 days
inclusive
60
First trimester ultrasound
Nuchal translucency (NT)
11–14
80
NT plus FTS
11–13
90
Second trimester Biochemistry
(HCG, oestridiol and alphafetoprotein)
14–18
60–70
Second trimester ultrasound
markers (e.g. echogenic cardiac foci,
pyelectasis, nuchal fold thickening)
18–20
20–30
(low sensitivity and
high FPR)
Reference : Australian Family Physician, Vol 32, No.3, March 2003, Advances in prenatal screening, pg 108
Primidone
Specimen:Plasma – Lithium heparin
Trough level is taken just before next dose (less than one hour).
Peak level is collected 1–3 hours post dose.
Reference Range: Supplied with report
Progesterone
Specimen:
Serum – Gel
Reference Ranges:Follicular < 1.0 mmol/L
Luteal 3.7–45 mmol/L
•
•
•
•
o detect ovulation a specimen is taken 6–9 days after presumed ovulation,
T
i.e. days 20–23 of a 28–day cycle. Serum levels rise sharply during the luteal
phase to reach a plateau.
Because hormone release is episodic, levels can vary considerably throughout
the day.
Pregnancy – levels rise steadily to reach a peak of 200–700 mmol/L by term.
Ectopic pregnancy – the diagnosis of ectopic pregnancy is supported by a level
below 15 mmol/L in the presence of symptoms.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Prolactin
Specimen:
Serum – Gel
Reference Ranges:Adult female
Adult males
< 540 mIU/L
< 390 mIU/L
Specimens should be collected at least 3 hours after awakening. Elevations of
up to 1000 can usually be ignored or even up to 2000 in normally menstruating
women. Repeat sampling 30 and 60 minutes after the first specimen will help show
up elevations due to stress alone. Prolactin levels above 5000 are almost always
associated with prolactinomas or pregnancy.
In women, hyperprolactinaemia is an important factor in infertility, amenorrhoea
and galactorrhoea (though 2/3 of women with galactorrhoea have a normal serum
prolactin).
In men, hyperprolactinaemia lowers testosterone levels and is an uncommon cause of
impotence or galactorrhoea as well as infertility.
Causes of hyperprolactinaemia
Physiological
• Stress
• Sleep
• Pregnancy – starts to rise at 6 weeks, peaks at up to 10,000 mU/L by term
• Lactation – peaks occur during episodes of breast–feeding with levels up to 5000
mU/L. prolactin is essential for normal lactation.
• Early morning collection – trough levels occur late morning
• Eating – small rises.
Drugs
• Androgens
• Cimetidine
• Haloperidol
• Methyldopa
• Metoclopramide
•
•
•
•
•
Oestrogens, including oral contraceptives
Opiates
Psychotropics
Reserpine
Tricyclic antidepressants.
Physiological
• Pituitary adenomas and micro–adenomas
• Other hypothalamic–pituitary disorders
• Polycystic ovary syndrome
• Hypothyroidism
• Renal failure
• Seizures
• Anorexia nervosa
• Addison’s disease
• Hypoglycaemia
• Idiopathic.
P
A
Prostate Specific Antigen (PSA)
Specimen:
Serum – Gel
Reference Range:Age related, total PSA
40–50 years
60 years 70 years 80 years < 2.6 µg/L
< 3.6 µg/L
< 4.6 µg/L
< 6.6 µg/L
Risk Assessment–Summary of Approach
Consider the age–related risk of prostate cancer
The prevalence of prostate cancer, as demonstrated in autopsy studies, is up to 80%
by the age of 80. The probability of clinical prostate cancer is under 1% for men at age
40 but rises progressively with age.
Consider the Total PSA
Assess PSA level according to age related reference values. A PSA greater than
20 µg/L is highly suggestive of cancer. If the PSA is less than 20ug/L but above normal
for age, check the Free/Total PSA value. Remember – a normal PSA does not exclude
cancer.
Consider the Free/Total PSA value
If the value is above 20% cancer is unlikely, though not excluded. If the value is less
than 8% there is a high probability of cancer.
On the basis of this approach definitive diagnosis by multiple prostate needle core
biopsies may be performed.
Further appropriate urology treatment and follow up can then occur.
Protein C
Specimen:Plasma – Sodium Citrate
Double spin, separate and freeze plasma.
Reference Range: Supplied with report
Deficiency of Protein C is an inherited cause of hypercoagulability and is found in
about 5% of cases of venous thromboembolism. The incidence of the deficiency is
1:5000–1:5,000. Approximately half of these will develop a thrombosis by the sixth
decade. Inherited deficiency is usually an indication for long–term anticoagulation.
Severe homozygous deficiency is associated with purpura fulminans in neonates.
Acquired causes of Protein C deficiency are liver disease, warfarin therapy,
vitamin K deficiency.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Protein S
Specimen:Plasma – Sodium Citrate
Double spin, separate and freeze plasma.
Reference Range: Supplied with report
Deficiency of Protein S, a cofactor for activated Protein C, is an inherited cause of
hypercoagulability and is found in up to 5% of patients with inherited thrombolic
disease. The lower end of the reference range is not well defined.
Acquired causes of Protein S deficiency are liver disease, warfarin therapy, vitamin K
deficiency, oral contraceptives, pregnancy.
Proteins, total, serum
Specimen:
Serum – Gel
Reference Range: Adults 60–80 g/L
Total proteins, consisting of albumin and globulins, is a crude test, but a raised
value can be a pointer towards raised immunoglobulins; a low value can be due to
reduced albumin or globulins or both. Variations in protein concentration can be due
to dehydration, diuretics, fluid retention or diurnal changes. On changing from the
recumbent to the upright position, fluid is redistributed to tissues from the circulation
causing an increase of up to 10% in protein concentrations.
Protein–binding in serum
Many serum constituents exist in two forms, a protein–bound form, usually to an alpha
or beta globulin or albumin; and a free form, often a tiny percentage of the total but
this is the metabolically active form.
Examples of protein–bound constituents are thyroxine, tri–iodothyronine, cortisol,
testosterone, vitamin B12, iron, copper, calcium–and many drugs.
Total globulins=total protein + albumin
P
A
Evaluation of Hyperglobulinaemia (Suggested scheme for evaluation of Hyperglobulinaemia)
Serum Protein Electrophoresis and Urinary Bence-Jones
Monoclonal Band
Benign
Present in 1–5% of subjects over 60 yrs
Paraprotein: <10g/L
Bence Jones: Negative
Serum Albumin: Normal
Other serum
Gamma–globulins: Normal
Haemoglobin: Normal
No increase over 3 year period
Malignant
Multiple myeloma
Plasmacytoma
Waldenstrom’s macroglobulinaemia
Malignant lymphoma
Chronic lymphatic leukaemia
Heavy chain disease
Amyloidosis (primary)
Polyclonal Hyperglobulinaemia
Chronic bacterial infection
Chronic parasitic infection
Autoimmune disorders
Chronic liver disease
Granuloma (e.g. Sarcoidosis)
Further investigations:
ANA ± ENA if autoimmune disease is
likely;
ACE (angiotensin converting enzyme) if
granulomatous disease is likely.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Proteins, urine (Proteinuria)
Specimen:
Random urine – Timed 10,12 or 24 hours
Reference Range: Supplied with report
Test methods
These depend on the clinical situation:
•
•
•
•
•
Urine dipstick, as part of routine urinalysis
Total protein concentration on spot urine – this gives a more accurate
measurement than a dipstick and also detects non–albumin proteins such as free
light chains.
24–hour protein quantitation–this test is used as follow – up when a dipstick shows
1+ (or more) positive, or when monitoring known proteinuria.
Urine microalbumin – a sensitive test measuring small quantities of albumin
in the range 0.01–0.20 g/L. The test is used as an early indication of diabetic
nephropathy.
Electrophoresis of concentrated urine – used mainly in detection of free light
chains (Bence Jones Proteins).
Renal proteinuria
Causes include the whole differential diagnosis of renal disease.
Glomerular proteinuria is by far the most common and serious type. The protein is
predominantly albumin and when daily output > 3g/day, nephrotic syndrome develops,
comprising oedema, albuminuria and hypoalbuminaemia. Serum lipids become elevated.
In tubular proteinuria, electrophoresis shows a non–selective pattern with bands
representing the wide range of normal serum proteins normally reabsorbed by the
tubules. Causes include drugs, inherited disease (e.g. Wilson’s disease), autoimmune
disease (e.g. SLE) or chronic infection.
Proteinuria of > 1.0 g/day requires renal investigation including consideration of renal
biopsy.
Benign and non–renal proteinuria
40% of our routine urinalyses show a trace or more of protein and most of these are
benign.
Causes include:
•
•
•
•
•
•
•
•
•
Infection
Fever
Stress
Exercise
Heart failure
Orthostatic proteinuria, found particularly in young men, disappears when the patient
is recumbent. Protein is absent from a morning specimen collected on first getting up.
Idiopathic
24–hour protein in these is usually < 0.5 g/day.
Levels of 0.2–0.5 g/day (or even up to 1 g/day) cannot be described as normal but
often remain of unknown aetiology when found as isolated abnormalities. Intensive
follow–up is usually unrewarding.
P
A
Prothrombin mutation (20210G–A)
Specimen:
Whole blood – EDTA
The prothrombin gene mutation 20210G–A, found in 1–3% of the population, is
associated with levels of prothrombin which exceed those in the normal population by
an average 30%.
Clinically the mutation is associated with:
•
•
•
•
Thrombophilia
3–6 fold increase in VTE
Cerebral vein thrombosis
Myocardial ischaemia in patients age < 50.
Pseudomonas aeruginosa
In general practice, Pseudomonas is encountered mainly in chronic otitis externa
where it usually responds to local toilet and topical polymyxin. The most commonly
used agent in the past, Chloromyxin, which contained polymyxin B, is no longer
available. Another ear preparation, Colymycin S Otic, containing colistin, which has
the same spectrum of antibacterial activity as polymyxin, is currently the most widely
used topical preparation for Pseudomonas otitis externa.
P. aeruginosa is often recovered from ulcer lesions on the lower limbs of the elderly. It
is frequently a coloniser rather than a pathogen in this situation. If, however, the gram–
stain reveals many WBCs with gram–negative bacilli and the ulcer has deteriorated
clinically Pseudomonas may be the cause of infection. In this instance ciprofloxacin
treatment may be indicated.
Pseudomonas can cause more serous infections in burns, in immunocompromised
patients and in nosocomial infections. In these situations, early treatment with carefully
selected antibiotics is required because of the organism’s pathogenicity and resistance
to a variety of agents.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Q
Q
A
Q Fever Antibodies
Specimen: Serum – Gel
Reference range: Supplied with report
Queensland Tick Typhus Antibodies
Specimen: Serum – Gel
Reference range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
R
R
A
RAST (Radioallergosorbent Test)
Specimen:
Serum – Gel
RAST tests use the patient’s serum to diagnose specific allergies by identifying
the presence of an IgE antibody against that allergen. Specify which allergens are
clinically relevant. A general Alatop screen can be performed, as well as a limited
number of individual allergens per test episode.
Red Cell Folate
Investigation of serum and red cell folate levels has been useful for the investigation
of macrocytosis, megaloblastic anaemia and nutritional status along with assessment
of vitamin B12 stores. Whilst red cell folate provides an accurate assessment of
tissue folate status, serum folate is of little diagnostic value since it can be reduced
with a short term decrease in dietary intake or in systemic illness. For these reasons
Capital Pathology will no longer be performing serum folate as part of B12 and folate
assessment. Red cell folate measurement will continue, and should now become the
main indicator for assessment of folate status.
Reducing Substances in Faeces
Specimen:Fresh sample of faeces delivered to lab as soon as possible,
stored in fridge if delay is unavoidable.
Reference Range: Not detected
The test is a simple qualitative test for reducing substances which include lactose,
glucose and fructose but not sucrose.
Acquired lactase deficiency, in which the disaccharide lactase is not digested,
presents as frothy diarrhoea with failure to thrive in infants after infectious diarrhoea.
The diagnosis can be confirmed by improvement after withdrawal of lactose from the
diet.
Renal Calculi
Most calculi are of unknown origin but four aetiologies need to be remembered:
1. Hypercalcaemia – check serum calcium, vitamin D
2. Uric acid excess in urine
3.Cystinuria
4.Mixed “triple–phosphate” stones, often due to chronic, occult urine infections with
gram–negative bacteria such as Proteus.
The reason for analysing calculi is to detect pure uric acid (5–10% of all stones) or
cystine. In the case of uric acid stones, the causes of increased uric acid excretion
need to be looked for. Allopurinol is effective in preventing further stone formation.
The remaining 90% of stones are composed of calcium oxalate or phosphate and in
half of these there is hypercalciuria due to increased intestinal absorption of calcium.
A 24–hour urine should be analysed for calcium, phosphate, urate and oxadate.
Renin
Specimen:Plasma – EDTA x 2
Spin,separate and freeze plasma within 2 hours of collection.
Reference Range: Supplied with report
Reticulocyte Count
Specimen:
Whole blood – EDTA
Reference Range: Adults absolute count 20–100x109/L
Absolute counts are preferred to percentages because they are independent of
anaemia. Polychromasia is the morphological indicator of increased reticulocytes.
The reticulocyte count is increased in:
•
•
•
•
Chronic or acute blood loss
Haemolytic anaemias
Deficiency anaemias treated with iron, B12 or folate (treatment response)
Marrow infiltration.
A reduced reticulocyte count suggests a hypoplastic basis for an anaemia.
Rheumatic Fever
Diagnosis is based on the revised Jones criteria:
Major manifestations
• Carditis
• Polyarthritis
• Chorea
• Erythema margination
• Subcutaneous nodular
Minor manifestations
• Clinical
– previous rheumatic fever
– or rheumatic heart disease
– or arthralgia
– fever
• Laboratory
– raised ESR, or CRP, or WBC
• ECG
– prolonged P–R interval.
Evidence of streptococcal infection
• Increased streptococcal antibodies
• Positive throat culture of GpA streptococcus
• Recent scarlet fever.
Capital Pathology Handbook – Interpretation of Laboratory Tests
The diagnosis requires:
• Two major criteria
or
•
One major and two minor criteria.
plus evidence of streptococcal infection
Absence of evidence of streptococcal infection makes the diagnosis doubtful. A serum
sample should be collected for streptococcal antibodies whenever the diagnosis or
rheumatic fever is suspected and a second sample collected at least two weeks later,
looking for rising titres.
SeeStreptococci
Rheumatoid Arthritis (RA)
The commonest of the connective tissue diseases with an incidence of 1–2% in the
general population. Patients may show:
•
•
•
•
•
•
•
Rheumatoid factor (positive in 80%)
Morning stiffness
Arthritis of hand joints
Symmetric arthritis
Arthritis of three or more joint areas (of 14)
Rheumatoid nodules
Radiographic changes.
Note that 20% of patients with rheumatoid arthritis are negative for rheumatoid factor.
See Rheumatoid Factor
Rheumatoid Factor (RF)
Specimen:
Serum – Gel
Reference Range: Supplied with report
Rheumatoid factor is an antibody, or rather a group of IgM, IgG and IgA antibodies,
directed against IgG. Most methods detect the same IgM rheumatoid factor but
specificities vary somewhat and for this reason a test that is positive in one laboratory
may be negative in another and the quantitations may differ.
2% of the healthy population are positive for RF, rising to 5–10% in the elderly.
In rheumatoid arthritis, RF is present in only 80% of cases. RF on its own does not
signify rheumatoid arthritis and conversely a clear clinical presentation of RA does not
require RF to establish the diagnosis. RF is often absent during the first 6 months of
disease. Higher levels tend to be associated with more aggressive disease.
Conditions, other than RA, with a high incidence of RF are:
•
•
•
•
•
Sjogrens (90%)
SLE (30%)
Other connective tissue disorders
Chronic inflammatory/infectious disorders
Malignancy.
R
A
Rickettsial Serology
Specimen:
Serum – Gel
Reference Range: Supplied with report
Ross River Virus Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Rotavirus
Specimen:
Fresh faeces
Rotavirus is the commonest cause of diarrhoea in young children – 50% of children
under five hospitalised for gastroenteritis are infected with rotavirus. Essentially all
children are infected by the age of three with the peak incidence between 6 and 24
months. Adult infections are much less frequent. There is a clear winter peak. Spread
is by the faecal–oral route. It has a short incubation period, 24–72 hours.
Symptoms may be severe but usually settle within 7 days with oral fluid and electrolyte
replacement.
Rubella Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Tests:IgG for immune status
IgM for current infection.
Immune Status
The person with an IgG antibody level > 10 u/ml, can be assumed to have immunity.
With levels between 5 and 10, immunity may be present.
Below 5, immunity is either non–existent or slight and revaccination may be indicated,
though not in pregnancy where immunisation should be delayed until after delivery.
It is essential that pregnant women found to be non–immune be vaccinated after
delivery.
Infection during pregnancy
Because 85% of mothers infected during the 1st trimester give birth to infants with
congenital defects (eyes, ears, heart, brain), termination of pregnancy must always be
discussed when infection has been established.
The mother who has had contact with suspected rubella and whose immune status
is susceptible or unknown, should immediately have blood collected for IgM and IgG
antibodies with at least one more specimen three weeks later.
Capital Pathology Handbook – Interpretation of Laboratory Tests
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Salicylate
Specimen:Plasma – Lithium Heparin
Optimal collection time is just prior to next dose.
Please supply time of last dose.
Reference Range: Supplied with report
Salmonella Antibodies (Widal Test)
Specimen:
Serum – Gel
A test of limited value in the diagnosis of typhoid fever, blood culture being the
preferred diagnostic method.
Interpretation:
O antigenstitre > 1:160 and rising sharply over 7–14 days suggests current
infection
H antigen
> 1:160 suggests immunity
Vi antigen
high titre sometimes indicates the carrier state
Titres can be nonspecifically elevated in diseases not caused by Salmonella and
vaccination can give positive titres.
Salmonella Culture
Specimen:Faeces for diarrhoea
Blood culture for suspected typhoid fever.
Enterocolitis
Salmonella species make up 15% of the pathogens cultured from faeces in this
laboratory, the main reservoir being domestic animals (including poultry and eggs) and
infected humans, both symptomatic and carriers. Species causing enterocolitis include
S. typhimurium, S. choleraesuis and S. enteritidis. In healthy persons the episode
usually resolves in 2–3 days. Patients with impaired defences may require antibiotic
treatment, usually with cotrimoxazole, amoxycillin or ciprofloxacin. The chronic carrier
state can be treated with ciprofloxacin.
Typhoid fever
During the early stages, diagnosis is by blood culture which should be done repeatedly
when clinical suspicion is strong. From the second week on, faeces cultures may be
positive.
Sarcoidosis
A systemic disorder of unknown cause characterised by non–caseating granulomata.
Diagnosis usually requires histological examination of lung, skin, lymph nodes or liver.
Serum ACE (Angiotensin Converting Enzymes) is elevated in 2/3 of patients but
the test is not specific enough to be of diagnostic value, nor does it have prognostic
significance.
Serum calcium may be elevated.
See Angiotensin Converting Enzyme (ACE)
Scabies
Caused by the mite Scarcoptes scabiei and transmitted by direct skin to skin contact.
Transfer from clothes is possible but only if worn by infected people immediately
beforehand.
Incubation period is 2–6 weeks without previous exposure but only 1–4 days in those
who have been infected before.
Burrows are characteristic in scabies. These consist of skin–covered ridges
0.5–1.0 cm in length, often with a small vesicle at the end.
The diagnosis is usually made clinically. Treatment is usually with permethrin 5%
cream. Gamma benzene hexachloride 1% cream or lotion, and crotamiton 10%
cream or lotion, are alternatives. Pruritis may persist for some time after successful
treatment.
Schistosomiasis
Specimen:Urine, full volume preferably collected between noon and 4pm
when there is peak egg excretion.
Serum for antibody tests.
Request examination for schistosome ova. Eggs may be found in urine and stool as
early as five weeks after infection. Patients with a low worm burden may have few or
no eggs in urine or stool.
Although over 200 million people in Africa, the Middle East, S. America and the
Caribbean are infected with these blood flukes, schistosomiasis is rare in travellers
to these areas. Infection requires skin contact with contaminated fresh water as in
swimming or wading barefoot in paddy fields, waterholes, local streams, etc. The
infective form penetrates human skin, passes through a migratory phase in the lung
and liver and then moves to its final habitat in the portal venous system (S. mansoni)
or urinary bladder venous plexus (S. haematobium). Adult male and females live for
5–10 years.
Many travellers have acquired infection from Lake Malawi in South East Africa. It is
stated in some travel guides that this lake is free of infection risk. This not the case
and travellers should be advised against swimming in this lake.
The usual problem is the traveller who requires reassurance. Examination of faeces
for the highly characteristic ova; urine for ova and red cells; serum for serology, and
perhaps liver function tests, are adequate for this purpose.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Where more serious investigation is required, up to six specimens of urine and faeces
should be examined. Further investigation might include appropriate tissue biopsies
(rectum, bladder, liver).
Positive serology indicates present or past infection. Persons with negative stool and/
or urine tests but positive serology require treatment to avoid the uncommon but
catastrophic spinal cord complication of transverse myelitis.
Treatment with praziquantel and follow–up are best managed by those with
experience treating schistosomiasis.
Scleroderma
Scleroderma Diffuse
Also called systemic sclerosis, this is a connective tissue disease characterised by
diffuse cutaneous and/or systemic fibrosis.
Raynaud’s phenomenon is present in 95% of cases. It has an incidence of about
1:50,000.
Tests that are often positive include:
•
•
•
ANA (95%) usually with a nucleolar pattern
RF (25%)
ENA anti Sci–70 (30%).
Scleroderma Limited
• Previously called CREST syndrome
• Calcinosis
• Raynaud’s phenomenon
• Esophageal dysmotility
• Sclerodactyly
• Telangiectasia.
Tests that are often positive include ANA (80%) usually centromere pattern.
Scoline (Suxamethonium) Sensitivity
This autosomal recessive disorder renders the carrier apnoeic for an abnormally long
period after administration of scoline. Family members of an affected individual should
be tested. The dibucaine number is an additional test used in identifying carriers.
SeeCholinesterase
Selenium
Specimen:Serum – Whole blood – Trace element tube.
24-hour urine (nil preservative).
Reference Range: Supplied with report
Selenium is an antioxidant and is an essential trace element in humans and animals,
entering the food chain through plants. In the Keshan province of China, an endemic
cardiomyopathy has been attributed to the region’s severe soil selenium deficiency.
Brazil nuts are a rich natural source of selenium.
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Seminal Fluid
1.
Post–vasectomy specimens
This is a test for sperm count only, not motility. The specimen should be delivered to
the laboratory on the day of collection but without the urgency necessary for fertility
specimens.
Most post–vasectomy specimens have a zero sperm count but a small percentage
have a low count of non–motile spermatozoa even months after successful surgery.
They are believed to be sequestered spermatozoa stored in the recesses of seminal
vesicles or other parts of the male GU system.
2.
Infertility specimens
The specimen should be delivered to the main laboratory within 2–3 hours of collection
and preferably before 3pm. Many people collect the specimen at home at 7–8am and
deliver the specimen before work. Please ensure specimen is kept warm. Patient
information sheets are available from the main laboratory, collection centres and the
website www.capitalpath.com.au
Serotonin (5–hydroxy tryptamine)
Specimen:Serum – Gel or
24-hour urine (HCL preservative).
Reference Range: Supplied with report
Sex Hormone Binding Globulin (SHBG)
Specimen:
Serum – Gel
Reference Range: Adult female
Adult male
20–155 nmol/L
15–100 nmol/L
SHBG is a serum globulin that binds testosterone and, to a lesser extent, oestradiol.
On its own, it has little diagnostic value but it is required when measuring the
biologically active free testosterone fraction as distinct from the 98% of total
testosterone which is bound and inactive.
SHBG is increased by: oestrogens, pregnancy, androgen deficiency, thyrotoxicosis
and cirrhosis.
It is decreased by: obesity, hypothyroidism, androgen excess, polycystic ovary
syndrome and hirsutism.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Sexually Transmitted Disease
1: Common presentations of sexually transmitted infections and recommended
investigations*
Presentation
Possible causes
Investigations*
Urethritis
(dysuria,
discharge,
frequency)
Common causes: Chlamydia
trachomatis (commonly
asymptomatic)
Neisseria gonorrhoeae
Trichomonas vaginalis
Mycoplasma genitalium, Mycoplasma
hominis, Ureaplasma urealyticum
(importance as pathogens not clear:
maybe associated with urethritis)
Herpes simplex virus (HSV) types
1 and 2 (can cause intermittent,
recurrent attacks of urethritis)
• First-catch urine (does not have to be first
void of the day) for PCR for C. trachomatis
(± N. gonorrhoeae)
• Urethral bacterial swab for microscopy and
culture (culture is necessary for antibiotic
susceptibility testing of N. gonorrhoeae)
• PCR is available and can also be done off
ThinPrep specimen
• Microscopy of wet preparation
• Urethral viral swab for HSV PCR.
Cervicitis
(usually
asymptomatic)
C. trachomatis
N. gonorrhoeae
• Urine, cervical or vaginal swab for PCR
for C. trachomatis and N. gonorrhoeae
• Cervical swab for microscopy and culture
for N. gonorrhoeae
Vaginal
discharge
(usually not
sexually
transmitted)
Candida albicans (most common
cause)
Bacterial vaginosis (caused by
changes in normal vaginal flora)
T. vaginalis (sexually transmitted)
• High vaginal swab for microscopy
and culture
Pelvic
inflammatory
disease
(pelvic pain,
menstrual
irregularities,
dyspareunia,
may be silent)
STI-related
C. trachomatis
N. gonorrhoeae
Non-STI-related
Mixed pathogens — anaerobes,
facultative bacteria and Mycoplasma
spp.
• Urine, cervical or vaginal swab for PCR for
C. trachomatis and N. gonorrhoeae
• Cervical swab for microscopy and culture
for N. gonorrhoeae
• Pelvic ultrasound examination
• Possible laparoscopy in severe cases
Genital warts
(typical warty
lesions in
anogenital
area)
Human papillomavirus (most
commonly types 6 and 11;
occasionally other types)
• Clinical diagnosis
Anogenital
ulceration
Infective causes
• Viral swab for viral culture ± PCR
for herpes
• Routine microsocopy and culture of swab
specimen (H. ducreyi is difficult to culture
on routine media)
• Serological tests for syphilis
• Biopsy for histopathological examination
• HSV-1 and HSV-2
• Syphilis (Treponema pallidum)
• Donovanosis
• Chancroid (Haemophilus ducreyi)
• Lymphogranuloma venereum
Non-infective causes
• Microscopy of wet preparation
• Biopsy if atypical lesions or doubt about
diagnosis
• Aphthous ulceration
• Behçet’s disease
• Trauma
• Drug reaction
• Inflammatory Bowel Disease
PCR = polymerase chain reaction. LCR = ligase chain reaction. STI = sexually transmitted infection. * Routine
screening for other STIs (including HIV) should also be offered to patients with any of these presentations.
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Shigella
Specimen:
Fresh faeces
Shigella, the cause of “bacillary dysentery”, is predominantly found in third world
countries where it is transmitted by contaminated food and water. Children under 5
years are particularly affected.
Mild cases are treated with oral rehydration alone but more severe infections may
require antibiotics and IV fluids. Shigellosis is a notifiable disease.
S. I. Units (Systeme Internationale)
In 1975–77, Australian, New Zealand and UK laboratories switched from traditional
mass units (mg/100ml) to SI units (mmol/L). United States laboratories have mostly
retained mass units which may need to be converted when American patients present
in Australia with results from back home.
SI Unit
Mass Unit
Conversion Factor
Alcohol
1 mmol/L
= 4.5 mg/100ml
0.22
Bilirubin
1 umol/L
= .06 mg/100ml
17.1
Calcium
1 mmol/L
= 4 mg/100ml
0.25
Cholesterol
1 mmol/L
= 38.5 mg/100ml
0.026
Glucose
1 mmol/L
= 17.9 mg/100ml
0.056
Potassium
1 mmol/L
= 1 meq/L
1.0
Protein
1 g/L
= .1 g/100ml
10.0
Sodium
1 mmol/L
= 1 meq/L
1.0
Triglyceride
1 mmol/L
= 91 mg/100ml
0.011
Urate
1 mmol/L
= 16.7 mg/100ml
0.060
Urea
1 mmol/L
= 5.9 mg/100ml
0.17
To convert mg/100 mL to mmol/L: multiply by conversion factor
To convert mmol/L to mg/100ml: divide by conversion factor
An advantage of SI units is that osmolality (in mmol/L) of serum or urine can be simply
calculated by adding concentrations (in mmol/L) of the constituents, assuming all the
main ones have been measured.
Sickle Cell Test
Specimen:
Whole blood – EDTA
Sickle cell haemoglobinopathies are hereditary disorders due to HbS, found in blacks
of African descent. The homozygous state (sickle cell disease) causes a chronic
haemolytic anaemia and during crises, episodes of pain and multisystem organ damage.
Heterozygotes (sickle cell trait) are free of clinical disease and have normal red cell
indices but HbS can be demonstrated by appropriate tests including Hb electrophoresis.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Sideroblastic Anaemia
This is an acquired disorder of porphyrin metabolism seen in older patients and
associated with anaemia, either microcytic or dimorphic. It is a preleukaemic condition
and is part of the myelodysplastic (MDS) spectrum of disorders. The hallmark is a
marked increase in bone marrow iron, ring sideroblasts or Perl stain in the bone
marrow and an elevated ferritin. The disorder may be primary (in MDS) or secondary
to drugs, particularly antituberculous therapy.
Sjogren’s Syndrome
A connective tissue disease characterised by salivary and lacrimal gland involvement
resulting in dry mouth and dry eyes.
It may exist on its own or be associated with rheumatoid arthritis or other connective
tissue disease.
Tests that are usually positive include:
•
•
•
ANA usually with a speckled pattern
RF (90%)
ENA (60%) anti SSA (Ro) or SSB (La).
Skin and Wound Swabs
When collecting the swab, aim to get pus or exudate from the base of an actively
inflamed lesion. Remove scab if present and open infected blisters. Avoid getting
bacteria from normal skin. 80% of skin/wound/pus swabs in the community grow
Staph. aureus. Strep. pyogenes makes up most of the remainder. The latter is the
usual pathogen in impetigo and cellulitis.
Chronic leg and foot ulcers are commonly associated with vascular insufficiency,
and, in the diabetic foot, with loss of pain and touch sensation. Swabs usually show a
mixed growth of gram–positive cocci and gram–negative bacilli. Cellutitis confined to
the rim of the ulcer can be treated with an oral agent such as amoxycillin–clavulanate
but a spreading cellulitis, particularly in a diabetic foot, needs urgent admission for
parenteral antibiotics.
All chronic ulcers require careful daily cleansing with warm saline and debridement of
dead tissue down to a healthy base.
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Skin Biopsy
Method
1. Excisional skin biopsy
This technique should be used for:
• Atypical pigmented lesions
• Deep dermal/subcutaneous nodules
• Where evaluation of margins is important
• Where panniculitis (inflammation of subcutaneous fat) is suspected.
2. Punch biopsy
This technique is useful for:
• Inflammatory dermatoses including alopecia
• Suspected basal or squamous cell carcinomas prior to definitive treatment.
Excisional biopsy should be performed rather than punch biopsy if malignant
melanoma is suspected. The punch biopsy should include subcutaneous fat.
Crush artefact, when forceps are used to extract the biopsy, should be avoided.
The specimen should be immediately placed in 10% formalin.
3. Shave biopsy
This usually results in epidermis only or epidermis and superficial dermis.
Shave biopsy should not be used if there is any suspicion of malignant melanoma.
In acral sites (soles of feet, palms or hands) usually only keratin is obtained in a
shave biopsy.
4.Curettage
This is the least satisfactory form of skin biopsy, as the tissue is typically fragmented
and may not be satisfactory for histology. Curettage is suitable for some superficial
lesions.
Site Selection
This is of critical importance particularly in:
Inflammatory dermatoses – biopsy should be from a fully developed lesion and if the
lesions are in different stages of evolution, multiple biopsies should be taken. Treated
areas should be avoided. For alopecia investigation two biopsies should be supplied
for vertical and horizontal sections.
Vesiculobullous lesions, ulcers, pustules – biopsies should be taken from very early
lesions and should include normal skin.
Direct immunofluoresence microscopy
Biopsies should be placed in transport medium (NOT in formalin). This may be
obtained from the lab by phoning 02 6285 9857. Alternatively the specimen may be
submitted to the laboratory fresh on damp saline–soaked gauze.
The specimen should arrive in in the laboratory within 2 hours of being taken.
SeeHistopathology
Capital Pathology Handbook – Interpretation of Laboratory Tests
Skin Antibodies
Specimen:
Serum – Gel
Reference Range: Not detected
Specify whether pemphigus or pemphigoid
Sodium, serum
Specimen:
Serum – Gel
Reference Range: Adults 135–145 mmol/L
Hyponatraemia
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Sodium, urine
Specimen:
Spot urine or 24-hour (nil preservative)
Reference Range: Supplied with report
In a well person, urine sodium output reflects dietary intake. A low output, < 20 mmol/
day, may be an indicator of a total sodium deficit but interpretation requires
consideration of all fluid and electrolyte parameters.
Sperm Antibodies
Specimen:
Semen sample and serum – Gel
Reference Range: Not detected
Antibodies in seminal fluid are a marker for immunological male infertility.
Spherocytes
Spherocytosis is associated with haemolysis:
•
•
•
•
•
Congenital spherocytosis
Autoimmune haemolytic anaemia
Microangiopathic haemolysis
Severe burns
After splenectomy.
Sputum Culture
Sputum is examined in three clinical situations:
1. Suspected TB
Where infectious TB is a possibility, ZN stain and culture are essential.
SeeTuberculosis
2.Pneumonia
If the patient can produce sputum – often a problem – the examination is worthwhile
provided it is collected before antibiotics are commenced.
3. Acute or chronic bronchitis
A sputum examination is seldom useful though the matter is debated. In exacerbations
of chronic bronchitis the important organisms are Strep. pneumoniae and
Haemophilus influenzae. Moraxella (formerly Branhamella) catarrhalis may also be
involved.
Sputum Cytology
Sputum cytology is useful for the investigation of bronchopulmonary and metastatic
tumours. Centrally located tumours have a higher rate of pick up on sputum cytology
than peripheral tumours.
Sputum cytology may also be useful for the investigation of asbestosis and various
infections, particularly in immunocompromised patients.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Patient Information for Collecting Sputum for Cytology
Routine sputum cytology series incorporates a separate specimen collected on each
of three consecutive days (the series is covered by one Medicare item number and
hence requires only one request form marked x 3).
Specimens should be collected early in the morning to maximise the yield of pooled
endobronchial material.
The mouth should be rinsed thoroughly with water prior to collection.
Deep cough specimens are important to ensure bronchopulmonary origin of the
specimen (oral and nasopharyngeal material are of little diagnostic value).
At least 2mL of sputum should be expectorated into a specimen container and
refrigerated pending delivery to the laboratory or nearest Collection Centre.
Each container needs to be labelled clearly with full name, collection date, date of
birth, the number in the series (i.e. 1, 2 or 3) and should be delivered to the nearest
Collection Centre as soon as practicable on the day it is collected. (Best results are
obtained on fresh specimens.)
Staphylococci
These organisms are divided into two groups according to whether they produce
coagulase, an enzyme–like protein that bestows invasive potential.
1. Coagulase-positive staphylococci
Staph. aureus
The common pyogenic organism of skin and wound infections
and a variety of serous systemic infections. About 30% of normal people carry Staph.
aureus in their anterior nares.
MRSA–methicillin resistant Staph. aureus
MRSA are always resistant to methicillin/oxacillin, penicillin, amoxycillin, amoxycillin–
clavulanate, and all cephalosporins.
With regard to other sensitivities, it is important to distinguish between two different
forms of MRSA:
Multi–resistant MRSA
Until 1995 these were the only variety known. They are resistant not only to penicillin
and cephalosporins but also alternative oral agents such as erythromycin, tetracycline
and cotrimoxazole.
They are found infrequently but when identified require action:
– Isolate the patient
– Minimise staff contact
– Screen staff if there is evidence of cross infection
– Treat infection
– Consider eradication therapy for those who are colonised
– Consult Infection Control Officer.
Non multi–resistant MRSA
In 1995 new strains of MRSA were identified that were seldom resistant to
erythromycin, tetracycline or cotrimoxazole. They do no require extensive infection
control procedures but should be treated with standard precautions as for other
S. aureus.
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2. Coagulase-negative staphylococci
Staph. saprophyticus
A relatively non–pathogenic organism. It is of clinical interest
mainly as a cause of urinary tract infections, particularly in young women where it is
second in frequency to E. coli.
Staph. epidermidis
A universal skin commensal, non–pathogenic except where
host defences are seriously compromised or when introduced by instruments or
prostheses.
S. epidermidis
Is not infrequently grown from an MSU but is of doubtful
pathogenicity. If accompanied by pyuria, treatment could be considered bearing in
mind the other causes of pyuria.
Streptococci
Streptococci causes a wide spectrum of common diseases. Three species or groups
are commonly isolated.
1.
Streptococcus pyogenes
Also known as beta–haemolytic group A streptococcus, this organism is the common
cause of bacterial pharyngitis. It also causes 20% of the skin and wound infections,
particularly cellulitis and impetigo where it is the most frequent pathogen. There are a
wide variety of other less common sites of infection.
2.
Streptococcus pneumoniae
Often called the pneumococcus, a common commensal in our upper respiratory tract,
with up to 60% of people carrying it. It is a gram–positive diplococcus which causes
alpha–haemolysis on blood agar (greening of the agar due to partial haemolysis of the
red blood cells) and often has a capsule conferring resistance to host defences. More
than 80 different serotypes are known but most disease is caused by a limited number
of serotypes. Transmission is from person to person by droplet spread.
It causes a wide range of infections: pneumonia with or without bacteraemia, otitis
media, sinusitis, meningitis, and other types of invasive disease. It does not cause
pharyngitis or tonsillitis. 30–50% of community–acquired pneumonia is due to
pneumococci and around 10% of nosocomial pneumonia.
Several conditions are associated with more frequent and severe pneumococcal
pneumonia: alcoholism, diabetes, chronic renal disease, heart failure and some
malignancies.
In the eye, S. pneumoniae is susceptible to chloramphenicol ointment but is
intrinsically resistant to neomycin.
Pneumococcal vaccine
Splenectomised patients are at significant risk for invasive pneumococcal disease and
should receive vaccine.
Pneumococcal vaccine contains 23 of the most common serotypes and covers 90%
of strains causing invasive diseases. 90% of adults respond to a single dose. The
vaccine is also recommended for persons > 65 years, persons at increased risk
of complications such as those mentioned above, immunocompromised patients,
including those with HIV
3.
Streptococcus faecalis (Group D streptococcus)
Capital Pathology Handbook – Interpretation of Laboratory Tests
Strongyloidiasis
Intestinal infestation with Strongyloides stercoralis is found worldwide in tropical
regions, particularly SE Asia and Central America. Clinically it can present with
abdominal discomfort, diarrhoea or malabsorption. A life–threatening disseminated
hyperinfection occurs in immunocompromised patients. Diagnosis relies on finding
larvae in faeces or, in disseminated disease, respiratory secretions. It tends to be less
easy to eradicate than hookworm which is a related nematode.
Synacthen Stimulation Test
Specimen:Serum – Clot or gel
Collect baseline specimen for cortisol.
Inject 0.25mg Synacthen (synthetic ACTH) IM.
Collect specimens 1/2 hour and 1 hour after Synacthen injection.
Indications:
Used in suspected Addison’s disease or for assessment of adrenal reserve after long
term steroid therapy.
Precaution:
Synacthen can cause a severe anaphylactic shock reaction leading to profound
shock and collapse. Test should only be performed by a clinician with access to full
resuscitation facilities. Test not performed by pathology laboratory or on outpatients.
Synovial Aspirate
Specimen:Collect into a plain tube
If sufficient fluid is available, collect a sodium citrate or EDTA.
Synovial fluid examination provides definitive diagnosis of septic arthritis, gout and
pseudogout, and places other effusions into the broad categories of non–inflammatory
lesions (trauma, osteoarthritis, etc.) and inflammatory non–infective disease
(rheumatoid arthritis, etc.).
Examination covers the following:
•
•
•
•
•
•
•
Gross appearance – clear, cloudy, purulent, blood–stained (trauma)
Volume
White cell count
White cell differential–polymorphs predominate in septic arthritis, gout,
pseudogout and sometimes in rheumatoid arthritis; mononuclear cells usually
predominate in the inflammatory non–infective disorders.
Gram stain
Crystals – polarised light is used to look for the urate crystals of gout or the
pyrophosphate crystals of pseudogout.
Culture – aerobes, gonococci and anaerobes.
Other examinations will be performed on request.
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Syphilis
Where there is a possibility of syphilis, serological tests should be performed. If a
lesion is strongly suspicious of syphilis with the typical painless indurated ulcer, direct
fluorescent stain can be arranged with the pathologist.
Specimen:
Serum – Gel
Reference Range: Non–reactive
1.
Screen test
The initial enzyme–immunoassay test is a screen for both non–treponemal and
treponemal antibodies. A positive screen test will be followed by the RPR (Rapid
Plasma Reagin), TPHA (Treponema Pallidum Haemagglutination) and FTA
(Fluorescent Treponemal Antibody).
2.
Non–treponemal (reagin) tests, RPR and VDRL
The RPR and VDRL (Venereal Disease Research Lab) are the most widely used of
the older, nonspecific reagin tests. They can be transiently positive in a wide range
of viral and autoimmune diseases as well as syphilis and yaws. After successful
treatment of syphilis, the RPR/VDRL titre declines and may become negative.
The RPR/VDRL is used as a screening test for syphilis and for monitoring response to
treatment. A positive result may be due to current infection.
3.
Treponemal tests TPHA and FTA
The TPHA and FTA are much more specific for treponemal infections but unexplained,
weakly reactive, false positives of one or both tests are sometimes found.
After a treponemal infection, the TPHA and FTA stay positive indefinitely without
indicating whether disease is current or past.
In true treponemal infections, all three tests, VDRL or RPR, TPHA and FTA, will
usually be clearly reactive.
Antibody tests cannot distinguish between yaws and syphilis infections which are both
due to Treponema pallidum species.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Systemic Lupus Erythermatosus (SLE)
A relatively common (1:1000) connective tissue disease which can affect a wide
variety of systems and is characterised by autoantibodies directed against nuclear
components. For initial investigation (screen), the following tests are recommended:
ANA, Anti- DNA, Lupus inhibitor, Rheumatoid Factor (RF).
There is no single specific test but rather a list of diagnostic criteria. The American
Rheumatology Association (1982) recommends that if four of the following criteria are
positive, the diagnosis can be made:
•
•
•
•
•
•
•
•
ANA positive (95%)
Anti–DNA, or ENA Sm, or biological false positive VDRL
Haemolytic anaemia, or leukopenia, or thrombocytopenia
Nephropathy (proteinuria > 0.5 g/day)
Arthritis
Pleuritis or pericarditis
Malar rash
CNS involvement (psychosis, seizures).
During active SLE, complement (C3 and C4) are reduced and the ESR is raised.
SeeAntinuclear Antibodies (ANA)
Extractable Nuclear Antigens (ENA)
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Capital Pathology Handbook – Interpretation of Laboratory Tests
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T3, free (Tri–iodothyronine)
Specimen:
Serum – Gel
Reference Range: 1.6–6.5 pmol/L
T3 is the active thyroid hormone with T4 being effectively a “prohormone”. 80% of T3
is formed from T4 in the tissues, the remainder being directly secreted by the thyroid.
Like T4, T3 is protein–bound in the blood and protein–binding abnormalities can
elevate both free and total T3 without causing hyperthyroidism.
Elevated by:
• Hyperthyroidism, including T3 thyrotoxicosis
• T3 therapy – because T3 has a short half–life, levels fluctuate up to 30%,
depending on time of last dose
• Subacute thyroiditis – early stage
• Hashimoto’s thyroiditis – occasionally in early stage
• Heterophilic antibodies, protein – binding abnormality.
Lowered by:
• Hypothyroidism – but is a poor test for this
• T4 therapy – though T3 is usually within the reference range when T4 dose
is appropriate
• Non–thyroidal illness
• Drugs: amiodarone, propranolol, steroids, lithium, iodine in tonics
or contrast medium
• Hypopituitarism (secondary hypothyroidism).
T4, free (Thyroxine)
Specimen:
Serum – Gel
Reference Range: 9.6–23.4 pmol/L
Although T4 is the principal thyroid hormone, it is converted to T3 in the tissues. It is
the preferred thyroid replacement therapy in hypothyroidism.
Elevated by:
• Hyperthyroidism
• T4 therapy–because the half–life is long, time of sampling for monitoring is
unimportant
• Non–thyroidal illness
• Drugs: amiodarone, NSAIDs, propranolol, steroids, iodine–containing contrast
medium, heparin
• Heterophilic antibodies, protein–binding abnormalities
• Sub–acute thyroiditis – early stage
• Hashimoto’s thyroiditis – early stage
• Self–administration of T4 e.g. for attempted weight reduction.
Lowered by:
• Primary hypothyroidism
• Non–thyroidal illness (uncommon)
• Hypopituitarism (secondary hypothyroidism)
• Hashimoto’s thyroiditis
• Sub–acute thyroiditis (recovery stage)
• Drugs: T3 (Tertroxin), phenytoin, lithium, carbamazepine.
Tegretol (Carbamazepine)
Specimen:
Serum – Gel
Trough level should be taken just before next dose (within one hour).
Peak level should be collected 3 hours post dose.
Therapeutic range: 15–40 umol/L
Testosterone
Specimen:
Serum – Gel
Reference Range:F <4.6 nmol/L
M 8 yrs < 2.5 nmol/L
M 14 yrs 3.0–10.0 nmol/L
M Adult 8.0–38.0 nmol/L
Free and total testosterone
Free testosterone, the biologically active fraction, is derived from total testosterone
using the value for SHBG which is the serum binding protein. Where the increase
in testosterone is small, as in minor degrees of hirsutism in the polycystic ovary
syndrome, it may be found that only the free testosterone is above its reference limit.
Age changes in males
The male embryo has high levels of testosterone falling to female levels at time of birth
but with a second peak during the first few months of life. Throughout childhood male
Capital Pathology Handbook – Interpretation of Laboratory Tests
levels are not much higher than in females. Until at about age 11 the pubertal rise
commences, reaching adult levels at about age 17. Levels decline slowly from about
age 40 on, the decline being more marked in free testosterone than total testosterone,
which is partially sustained by a rise in SHBG.
Thalassaemias
The thalassaemias are common hereditary conditions in which there is a reduction
in the synthesis of one or more of the four globin chains of the Hb molecule. Adult
haemoglobin, HbA, which makes up > 96% of normal Hb, contains two alpha and two
beta globin subunits and these define the two main groups of disorders, the alpha
thalassaemias and the beta thalassaemias.
The genes for thalassaemia are found commonly in Asians, Polynesians, Africans
and Mediterranean people. Clinically, the thalassaemias range from the clinically
undetectable heterozygous state, through mild microcytic, hypochromic anaemias,
(thalassaemia minor) to severe transfusion–dependent anaemias or fetal death
(thalassaemia major).
Clinical Presentation
The major thalassaemia syndromes usually come under specialist care early in life
with moderate or severe haemolytic anaemias.
•
•
•
•
•
he thalassaemia trait disorders, which are usually asymptomatic, will for the most
T
part be discovered incidentally on a routine blood film.
Anaemia, usually mild, sometimes moderate, sometimes lower end of the
reference range
Microcytic, hypochromic blood film, resembling an iron deficiency anaemia but
with an MCV which is disproportionately low compared with the anaemia
Normal ferritin, s. iron, iron–binding capacity
The blood film may show other suspicious abnormalities; and the histogram of red
cell size distribution may show a pattern typical of thalassaemia.
Diagnosing the Type of Thalassaemia
The thalassaemias are complex and diverse and under certain circumstances may
require specialised DNA and globin chain analyses to characterise them fully. In
ordinary clinical practice, once the diagnosis of thalassaemia has been suspected
because of microcytosis in the absence of iron deficiency, a group of tests is applied to
broadly separate alpha from beta thalassaemias.
These tests are:
• HbH inclusion bodies (alpha)
• Hb electrophoresis
• HbA2 % (beta)
• HbF % (beta)
• Kleihauer stain (beta).
T
A
Alpha Thalassaemias
Each parent contributes two alpha genes giving a total of four. Alpha thalassaemia
results from deletion of one or more of these four genes.
Deletion
Hb
MCV
HbH bodies
clinical
– alpha/alpha
alpha
single
gene
N
N
Absent
‘silent carrier’
–alpha/–alpha
two
gene
(trans)
N
N (usually)
Very
occasional
Polynesian
type
–/alpha alpha
two
gene
(cis)
±/

Occasional
more severe
than trans.
asian type
–/–alpha
three
gene


Numerous
HbH disease
–/–
four
gene
–
–
–
Intrauterine
death
Beta Thalassaemias
Beta thalassaemias arise from point mutations in the coding genes rather than
deletions and more than 300 mutations have been identified. The first thalassaemia to
be described, by Cooley in 1925, was severe beta thalassaemia found in a patient of
Mediterranean descent, thalassa being the Greek for sea.
Alpha and beta thalassaemias occur with roughly equal frequency.
Theophylline (Nuelin)
Specimen:
Serum – Gel
Therapeutic range: 55–110 µmol/L
The therapeutic range refers to peak levels. The specimen is collected 4–6 hours after
the last dose for long–acting preparation, 2 hours after those that are short–acting.
ThinPrep
See
Cervical Cytology
Throat Swabs
Swabs are taken from the tonsillar area for a sore throat, or the posterior pharyngeal
wall for sinus trouble so as to collect post–nasal discharge.
Capital Pathology Handbook – Interpretation of Laboratory Tests
The most common indication for a throat swab is to detect Group A streptococcus
(S. pyogenes). Culture results take 1–2 days to be reported.
Rapid streptococcal antigen tests are not performed in the laboratory. Their general
use increases the cost of testing as it is commonly recommended that all patients with
a negative rapid antigen test require a culture.
Streptococcus pyogenes, which is 100% sensitive to penicillin, is by far the
commonest bacterial pathogen in pharyngitis in immune competent patients.
Occasional pathogens include Arcanobacterium haemolyticum, Bordetalla, gonococci,
groups C and G streptococci, diphtheria, and anaerobic organisms in quinsy.
Thrombocytopenia
A reduction in platelets below 150x109/L. The lower the platelet count, the stronger the
possibility of spontaneous bleeding.
0–20x109/LSevere thrombocytopenia which can be life–threatening.
Should always be referred to a haematologist.
Bleeding is not directly proportional to the degree of thrombocytopenia.
Thrombocytopenia in pregnancy can put the fetus at risk and should be referred for a
specialist opinion.
Causes of thrombocytopenia:
Acute infection
Transient, often marked, thrombocytopenia may be seen in association with
acute viral illnesses in children. In this setting, platelet levels often recover rapidly.
Post–viral thrombocytopenia in adults may persist at mild to moderate levels and
is assumed to have an immune–mediated mechanism. Acute HIV infection may be
a cause of thrombocytopenia.
Drugs
A long list including salicylates, sulphonamides, trimethoprim, penicillins,
cephalosporins, methyldopa, chlorthiazide, frusemide, tolbutamide, heparin,
phenytoin, phenobarbitone, carbamazepine, phenothiazines, phenylbutazone,
gold, penicillamine – and others.
Alcohol and liver disease
ITP
Idiopathic Immune Thrombocytopenic Purpura
Leukaemias
Marrow infiltration
Malignancy, myelofibrosis
Other
Hypersplenism, SLE, B12 or folate deficiency, DIC, post–transfusion, post–partum
Pregnancy
ITP, dilutional or gestational thrombocytopenia, GPH syndromes
T
A
Thrombocytosis
Refers to an increase in the platelet count above 450x109/L.
Transient reactive thrombocytoses up to about 800x109/L are common. Causes are:
•
•
•
•
•
•
•
•
•
•
Blood loss
Surgery, trauma
Infection, including viral infection
Other inflammatory disorders
Malignancy – an important cause of persistent thrombocytosis
Myeloproliferative disorders
Essential thrombocythaemias
Polycythaemia vera
Myelofibrosis
Myelodysplasia.
Thrombophilia
Patients with thrombosis may be further investigated, especially those with thrombosis
at a younger age, thrombosis at an unusual site, those with a family history of
thrombosis or those with recurrent thrombosis.
Non-genetic risk factors for thrombosis must also be assessed for cumulative risk
assessment such as hypertension, smoking, diabetes mellitus, obesity and the oral
contraceptive .
Tests may include:
•
•
•
•
•
•
•
•
•
ntithrombin III (ATIII)
A
Protein C
Protein S
Lupus Inhibitor
Anticardiolipin Antibodies
Homocysteine levels
Activated Protein C Resistance
Factor V Leiden Mutation
Prothrombin Gene Mutation.
Medicare benefit is available where the request for testing specifically identifies that
the patient has a history of venous thrombosis or pulmonary embolism or is a first
degree relative of a person who has a proven diagnosed defect.
Please specify individual tests requested.
Capital Pathology Handbook – Interpretation of Laboratory Tests
Thyroid antibodies
Specimen:
Serum – Gel
Please specify which thyroid antibody is required.
Reference Range: Not present
Graves’ disease and primary hypothyroidism are both autoimmune diseases and are
associated with a variety of antibodies.
Anti-TPO and anti-thyroglobulin antibodies
Elevated in:
•
•
•
•
ashimoto’s thyroiditis – antibodies in high titre will differentiate non–toxic goitre
H
from Hashimoto’s
Primary hypothyroidism – antibodies present in 90%, often in high titre
Autoimmune disease marker – there is an association with diabetes and
pernicious anaemia
Euthyroid normals – 10% have antibodies usually in low titre. Annual follow–up will
show progression to thyroid disease in some, indicating that these elevations can
be a marker for an early autoimmune state.
Thyroid Stimulating Antibody (TS antibody, also called TSH receptor antibody)
This IgG antibody, formerly known as LATS, is the marker for autoimmune
thyrotoxicosis (Graves’ disease).
Thyroid Stimulating Hormone (TSH)
Specimen:Serum – Gel
Please specify which thyroid antibody is required.
Reference Range: Adults 0.50–6.3 mIU/L
Abnormal TSH levels, whether high in hypothyroidism or low in hyperthyroidism,
respond slowly to appropriate therapy with the new equilibrium level not reached for
2–8 weeks.
TSH is elevated by:
• Primary hypothyroidism, subclinical or clinical
• Hashimoto’s thyroiditis
• Subacute thyroiditis, recovery phase
• Non–thyroidal illness – occasionally during recovery phase
• Ectopic TSH from tumours of lung, breast etc.
• Drugs: lithium, metoclopramide, clomiphene, domperidone, iodides: kelp tablets,
amiodarone, contrast medium
• TSH release is pulsatile – minor elevations may simply be detecting the transient
peak of a pulse.
TSH is decreased by:
•
•
•
•
•
yperthyroidism – usually the TSH is < 0.03 in clinical thyrotoxicosis
H
Thyroid autonomy/sub–clinical hyperthyroidism
Patients on supra–optimal T4 or T3 therapy
Drugs: steroids, L–dopa, bromocriptine, heparin
Non–thyroidal illness.
T
A
Suppressed TSH
Elevated TSH
Capital Pathology Handbook – Interpretation of Laboratory Tests
Tissue Typing
Specimen depends on the history – Whole blood ACD tube, clot tube, EDTA plasma or
SST tube. Keep specimen at room temperature.
Can be collected on Mondays to Thursdays, however, a booking needs to be made
with Red Cross by the doctor or Collection Centre prior to specimen collection.
Red Cross: 02 9234 2332
TORCH Screen Antibodies
This is an acronym derived from infections causing intrauterine growth retardation or
death: Toxoplasma, Other, Rubella, CMV, Herpes (and Syphilis).
“TORCH antibodies” is no longer regarded as an appropriate block of tests –
individual tests should be requested according to indications.
Toxoplasma Antibodies
Specimen:
Serum – Gel
IgM antibodies become detectable 5 days after infection and remain for months or
occasionally years. A positive IgM result does not separate current from past infection
except when a rising titre can be demonstrated. Approximately 2% of women tested
antenatally are +ve for IgM but most do not have active infection.
IgG antibodies become positive 1–2 weeks after infection and remain positive for
life. A strongly rising titre over a 3–week interval is good evidence of current infection.
30–60% of the population have IgG antibodies.
Life–cycle and clinical disease
Toxoplasma gondii is an intracellular protozoan found in cats, humans, sheep, pigs
and other mammals. Cats are the primary hosts spreading cysts in their faeces to be
accidentally ingested by cat–lovers and grass–eating animals. In the secondary host
(man, domestic animal) the infection is usually subclinical but with lymphadenopathy,
variant lymphocytes in the blood film and a lymphocytosis. Viable parasites in the
tissues of domestic animals cause infection when their meat is eaten undercooked.
In the immunocompromised human, quiescent lesions can be reactivated causing
serious disseminated infections.
Toxoplasmosis in pregnancy
About one third of women acquiring toxoplasmosis during pregnancy will transmit
the parasite to the fetus. In the first trimester the incidence of infection is about
10%, but with a high risk of serious or fatal disease in the fetus. In the second and
third trimesters the fetal infection rate rises to 30% and 60%, respectively, but with
less serious effects in the fetus where the disease may not be apparent until later in
childhood with CNS impairment or chorioretinitis.
If antibodies were known to be present at least one month before conception, the fetus
will be safe.
Because infection, whether in or out of pregnancy, is usually subclinical, the diagnosis
is often made only when an affected fetus or child is encountered.
Sometimes a mother will ask for toxoplasma tests during pregnancy and about 2%
of these will test +ve for IgM antibodies, most of them derived from pre–pregnancy
infections.
T
A
If a mononucleosis–like illness occurs in pregnancy, or if there is any other reason to
suspect acute maternal infection, serial testing of antibodies is obligatory followed by
toxoplasma DNA testing of amniotic fluid or fetal blood. DNA testing of amniotic fluid at
around 18 weeks of pregnancy is highly predictive of the presence or absence of fetal
infection. Discussion with a microbiologist is recommended when toxoplasmosis in
pregnancy is suspected.
Transferrin
Specimen:
Serum – Gel
Reference Range: 2.0–3.2 g/L
Transferrin is the serum iron–binding transport protein which is measured by “iron–
binding capacity” and is described under that heading.
As an approximation, transferrin = IBC ÷20.
Trichinosis
An intestinal and muscular infection caused by the nematode Trichinella spiralis. Pigs
are the main reservoir, and eating under–cooked pork the principal mode of infection.
Although T. spiralis is common worldwide, particularly in North America and Europe.
Australia is considered essentially free of infection. Infected pigs probably still exist. It
is likely that rats and wild cats serve as a reservoir.
Diagnostic tests include eosinophil count, CK, serological tests and muscle biopsy.
Triglyceride
Specimen:
Fasting Serum – Gel
Desirable Range: 0.5–2.0 mmol/L (fasting)
Triglyceride in the fasting state comes from the liver in VLDL particles and their smaller
IDL products. When triglyceride metabolism is markedly impaired, chylomicrons
can be present in the fasting state. If the fasting triglyceride is above 5 mmol/L, the
specimen will appear cloudy due to raised VLDL and/or chylomicrons which rise to
form a creamy layer on standing.
Elevated fasting triglyceride
1.Primary
• Familial combined hyperlipidaemia
• Familial hypertriglyceridaemia
• Type III (“remnant removal disease”) hyperlipoproteinaemia.
2.Secondary
• Obesity
• Alcohol
• Diabetes
• Hypothyroidism
• Liver disease, particularly obstructive
• Nephrotic syndrome
• Pancreatitis
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
•
•
Pregnancy
Significant illness
Drugs: oestrogen, oral contraceptives, beta blockers, corticosteroids, thiazides,
retinoic acid, antiviral agents, valproic acid.
Very high triglycerides
A patient with a triglyceride above 10.0 mmol/L is at risk for acute pancreatitis and
requires immediate restriction of dietary fat and alcohol, treatment of any other
underlying cause such as diabetes, and addition of a triglyceride–lowering drug such
as a fibrate if other measures fail.
With massive hypertriglyceridaemias, serum can have the appearance and
consistency of cream. Often there is more than one aetiology, e.g. diabetes and
alcohol.
Triglyceride levels can rise and fall very quickly. Above a level of 6.0 mmol/L,
lipoprotein lipase clearance mechanisms are saturated which means that dietary fat
can rapidly raise triglyceride to surprising levels. Dietary restrictions may cause it to
fall equally quickly.
See
Lipid Disorders
Troponin
Specimen:Canberra + Cooma region: Serum – Gel
Goulburn + Bega region: Whole Blood – Lithium heparin
Reference Range:Troponin T < 0.03 ng/mL
Troponin I < 0.03 ng/mL
The markers of myocardial damage are, in decreasing order of specificity, troponins I
& T, CK–MB, total CK, myoglobin, AST and LD.
AST and LD no longer have a useful role.
Myoglobin, though relatively nonspecific, is the earliest to rise and can appear within
the first six hours.
The remaining four typically start to rise 4–12 hours after infarction, reach a peak and
return to baseline over a period which differs between the four.
Elevated for
CK–MB
2–3 days
Total CK
2–3 days
Troponin I
4–7 days
Troponin T
4–10 days
T
A
Of these, the troponins are the most specific and usually, when elevated, indicate
infarction. Small elevations found in unstable angina may indicate ischaemic damage
and carry a poorer prognosis than a normal level.
Troponins do not rise earlier than CK but their longer period of elevation is useful, e.g.
when chest pain occurred several days earlier.
CK–MB is falling into disuse since the troponins arrived, though it may be useful in
plotting successive reinfarctions.
An elevated total CK is nonspecific but if serial CK measurements follow the typical
time course of an MI, they support the diagnosis. A small MI may occasionally be
suggested by CK changes that stay within reference range but show a typical peak
over 2–3 days.
Elevation of cardiac troponin is more sensitive and specific for myocardial infarction
than CK–MB. Concentrations rise with 4–12 hours of commencement of cardiac pain
and remain elevated for up to 10–14 days.
Unstable angina can also cause elevations due to minimal myocardial damage not
detected by CK–MB. It is important to recognise that angina without myocardial
necrosis will not elevate troponins.
If the specimen was obtained less than 6 hours after commencement of chest pain,
a follow–up should be collected.
Tuberculosis (TB)
One third of the world’s population is infected with TB, the majority of them in the
developing world.
High risk groups are those who have arrived from Asia or the Pacific Islands in the
past few years and those with other family members who have been infected.
Infection is by inhalation of droplet nuclei. Local replication leads to acute disease in
about 5% of those infected. It is higher in infants and those more than 60 years old.
About 10% of infected people reactivate during their lifetime. This is more common
over the age of 50 and in men more than women.
Diagnosis
Specific diagnosis requires isolation of bacilli from an appropriate specimen.
The tuberculin test can be useful provided its limitations are recognised.
Specimens:
UrinesThe whole of an early morning specimen should be collected
on three different days. A special collection jar is provided and
each should be returned to the lab on the day of collection.
SputumZN stain and culture of sputum is the most effective test
for diagnosing infectious TB. Three specimens should be
examined, preferably early morning on separate days.
Automated liquid–based culture methods grow TB from most
smear–positive specimens within a week.
Bronchial washingsCollect into a sterile container.
TissueSpecimens for culture must be placed in sterile saline or
water, not in formalin.
See Mantoux Test
Capital Pathology Handbook – Interpretation of Laboratory Tests
Tumour Markers
Specimen:
Serum – Gel
The principal value of tumour markers is in monitoring patients with known malignant
disease. The lack of tissue specificity for many markers as well as the occurrence of
false positive and false negative results limits their use as a screening or diagnostic
test. Some of the markers applied to tumour surveillance are shown below.
Tumour Marker
Condition
Alpha–fetoprotein (AFP)
Testicular cancer (non–seminomatous)
Hepatocellular carcinoma
Gastrointestinal tract cancers with and without liver
metastases
CA 125
Ovarian carcinoma (non–mucinous) liver, pancreatic,
lung, colon, uterine, fallopian tube
Carcino Embryonic Antigen
(CEA)
Colorectal cancer
Pancreatic, breast, lung, small intestine, stomach,
ovaries
Prostate Specific Antigen (PSA)
Prostate cancer
Human Chorionic Gonadotropin
(HCG)
Choriocarcinoma
Hydatidiform mole
Trophoblastic diseases
Turner’s Syndrome
In Turner’s syndrome, phenotypic females have only one X chromosome rather than
the usual two. Clinical features include short stature, sexual infantilism and primary
amenorrhoea. Gonadal dysgenesis is indicated by raised FSH and LH and low
oestradiol.
Diagnosis is based on demonstration of the abnormal karyotype by the cytogenetics
laboratory.
T
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
U
U
A
Urate, serum
Specimen:
Serum – Gel
Reference Range: Supplied with report
Urate is a breakdown product of cell nuclei, one third from the diet, two thirds from
endogenous tissue catabolism. 20% of urate is excreted by the kidney, the remainder
in the gut.
Elevated Urate
Like cholesterol, baseline serum urate levels are genetically determined but elevated
by secondary factors:
•
•
•
•
•
•
•
•
•
•
•
•
High purine diet – liver, kidney, shellfish, fish roe, kina
Alcohol
Renal insufficiency
Drugs: diuretics, salicylates in low dose, steroids, chemotherapeutic agents,
niacin, ethambutol, pyrazinamide
Polycythaemia vera
Malignancies, particularly leukaemias or lymphomas being lysed by chemotherapy
Hypothyroidism
Rare enzyme defects – can present as gout or urate nephropathy in children,
young adults or pre–menopausal women
Down’s syndrome
Metabolic syndrome – hypertension, dyslipidaemia, diabetes, obesity
Pregnancy – compared with the non–pregnant base–line, levels are 20% lower
in the 1st trimester and 20% higher in the 3rd
Gout – an acute inflammatory arthritis precipitated by urate deposition in
synovial tissue and occurring mainly in middle–aged and older men and in
post-menopausal women.
Demonstration of urate crystals in aspirated synovial fluid establishes the diagnosis
beyond doubt.
Raised serum urate levels are contributory but not diagnostic. During an acute attack
of gout, serum urate may actually fall below the levels that will be found between
attacks.
Evaluation of Hyperuricaemia
Exclude:
Potentially correctable contributory factors:
• Obesity
• Alcohol
• Hypertriglyceridaemia
•Drugs (especially Thiazides, but also Diuretics, Salicylates [low dose], Nicotinic acid,
Pyrazinamide, Ethambutol, Cyclosporin)
• Hypertension
• Low fluid intake
Consider:
High Purine intake
Decreased renal excretion
Diet (meats, yeast products)
Primary
• Idiopathic
• Enzyme defects
•Syndrome–X, (insulin resistance;
dyslipidaemia [increased TG, low
HDL–chol]; obesity; hypertension;
hyperuricaemia)
• Idiopathic
Secondary
Secondary
•
•
•
•
•
•
•
• Renal failure
If renal failure is causing the
hyperuricaemia, serum creatinine will be
> 0.40 µmol/L, serum urate < 0.65 mmol/L
and the ratio of urine urate: creatinine will
be < 0.7. If hyperuricaemia is causing the
renal failure, then serum urate > 0.7 mmol/L
and urine urate: creatinine ratio > 0.7.
• Dehydration
• Diuretics
• Ketonaemia (starvation; diabetes mellitus
•Hyperlactataemia (alcohol; toxaemia or
pregnancy)
• Drugs
• Hyperparathyroidism
Increased Urate production
Primary
Blood dyscrasias
Infectious mononucleosis
Malignancy
Cytotoxic therapy
Psoriasis
Alcoholism
Prolonged exercise
Capital Pathology Handbook – Interpretation of Laboratory Tests
Urea
Specimen:
Serum – Gel
Reference Range: Adults 3.0–8.0 mmol/L
Elevated by:
•
•
•
•
•
•
•
Renal impairment
– b
ut serum creatinine is a better test being less subject
to other interferences
High protein diet – an important determinant
Catabolic states – any acute serious illness, particularly sepsis
Dehydration­– at low urine flow, there is increased tubular reabsorption
of urea
Bleeding into GI tract – can give elevations up to 15 mmol/L
Prostatic hypertrophy – or other post–renal obstruction
Drugs– steroids, diuretics.
As a routine measure of renal function, creatinine has almost entirely replaced urea.
Nephrologists, however, measure urea as well as creatinine in dialysis patients,
the preferred urea level being below 20–30 mmol/L.
An unexpected fall can be due to loss of albumin in dialysis fluid.
Unexpected rises in dialysis patients can be due to:
•
•
•
•
Dietary non–compliance, too much protein
Sepsis
Dehydration or catabolic illness
Steroid dose too high.
U
A
Urethral Swabs
Male
Where there is a discharge, swabs and gonococci culture should be collected by the
doctor at the time of examination. In the absence of a discharge, urethral swabs are
unlikely to grow a pathogen.
Female
The urethra can be swabbed for gonococci when looking for STD.
Urine Cytology
Urine cytology has a high degree of accuracy in high grade urothelial tumours and
carcinoma in–situ. However the test is less accurate with low–grade neoplasia
including papillomas. Infection, lithiasis and catheterisation may be difficult to
distinguish from low grade urothelial neoplasia on urine cytology.
Patient Information for Collecting Urine Series for Cytology
Three samples of urine to be collected preferably on three consecutive days. (The
series is covered by one Medicare item number and hence requires only one request
form marked x 3).
The second urine of the morning is the preferred specimen. It should be collected
each morning, and a portion of this sample placed in the “specimen container”
provided. The container should be at least half filled.
Each container needs to be labelled clearly with full name, collection date, date of
birth, the number in the series (i.e. 1, 2 or 3) and should be delivered to the nearest
Collection Centre as soon as practicable on the day it is collected. (Best results are
obtained on fresh specimens.)
If there is any delay in delivery the specimen should be refrigerated.
Urine, pigmented/colouration
Causes include:
Red urine
• Haematuria
• Phenolphthalein–containing purgatives, which are red when alkaline. Fresh urine
is acid but turns alkaline on standing
• Diet–beetroot, rhubarb turn red on standing, particularly with coexisting iron
deficiency
• Some porphyrias–turn red/brown on standing
• Haemoglobinuria
• Myoglobinuria.
Orange / brown urine
• Concentrated urine in febrile states may be described as abnormal by patients
• Bilirubin, or urobilinogen which turns brown on standing
• Drugs, e.g. methyldopa, de Witts pills, chloroquine, rifampicin, quinine,
metronidazole, nitrofurantoin, riboflavin
• Some porphyrias turn brown on standing
• Melanin in disseminated melanoma
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
•
Alkaptonuria, turns brown on standing
Tyrosinaemia.
Yellow urine
• Vitamin B complex, multivitamins.
Blue / green urine
• Drugs, e.g. amitriptyline, NSAIDs, triamterene
• Biliverdin.
Milky urine
• Infection
• Chyluria
• Nephrotic syndrome
• Oxaluria.
Factitious additives (something the patient has added) are another source of
pigmentation.
Urine, 24–hour collection
Some tests require preservatives. Refer to specific test for further information.
A carefully timed specimen should be collected as follows:
1.The bladder must be emptied at a set time on day one (e.g. 8.00 am) and this
urine discarded.
2.All urine voided during the next 24 hours must be collected and transferred
into the large 24–hour urine container provided. The container should be kept
refrigerated.
3.The bladder must be emptied at the same time on day two as for the
commencement of the test on day one (e.g. 8.00 am). This specimen must be
included in the 24–hour specimen for assay.
4.The patient’s name and the dates and times of commencement and completion
of collection are to be noted on the container. If for Creatinine Clearance, include
height and weight of patient and a specimen for Serum Creatinine.
5.The specimen should be forwarded to the laboratory as soon as possible after
collection is completed.
Patient information leaflets are available from Collections Centres,
website www.capitalpath.com.au or front reception on 02 6285 9800
U
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
V
V
A
Vaginal Swabs for Discharge
Specimen:Routine bacterial swab collected from 5 cm inside the vagina and
placed in transport medium.
The normal flora of the vagina is an abundant growth of gram–positive lactobacilli.
Pathogens are of three main types:
•
Trichomonas
– detected by microscopic examination of a smear taken
from the swab
•
Candida
– often obvious clinically as a curd–like growth,
it is easily cultured
•
Bacterial vaginosis
– a
growth of predominantly anaerobic organisms
associated with a lack of normal lactobacilli. Clue cells
are a feature and Gardnerella vaginalis may be present.
Metronidazole provides effective treatment for both bacterial vaginosis and
Trichomonas infection.
Where an STD is suspected as cause of a discharge, cervical and urethral swabs
should be collected. Vaginal swabs seldom grow gonococci or test positive for
Chlamydia.
Valium (Diazepam)
Specimen:Plasma – Lithium heparin
Trough level suggested, taken before next dose (within one hour).
Reference Range: Supplied with report
See
Diazepam
Valproic Acid (Epilim)
Specimen:Serum – Gel
Suggest trough level collected just before next dose.
If peak requested collect between 0.5–1.0 hours for Syrup,
1–3 hours for capsules and 2–6 hours for coated tablets.
Reference Range: Therapeutic 350–700 µmol/L
SeeAnticonvulsants
Vancomycin
Specimen:Serum – Gel
Trough level is taken before next dose (within one hour).
Peak level is collected 30 min post completion of IV dose,
or one hour post IM injection.
Trough levels are preferred, trough should be less than 15 mg/L.
Reference Range:Pre dose trough 5.0–15.0 mg/L
Post dose peak 25.0–40.0 mg/L
Varicella–Zoster Virus (VZV)
Specimen:Viral swab taken from base of fresh vesicle or serum gel
for serology.
The same virus is responsible for both chickenpox (varicella) and shingles (herpes
zoster) the latter being a reactivation of dormant virus acquired during a childhood
attack of chickenpox.
Diagnosis is clinically obvious in most cases but the virus can be identified by PCR, a
fluorescent antibody applied to a suitable smear or by culture. PCR is the preferred method.
Venous Thromboembolism (VTE)
In hospital practice venous thromboembolism is thought to contribute to 10% of
deaths. In the community, incidence varies, depending on age, between 0.1 and
1/1000 per year.
Objective diagnoses of DVT (Deep Vein Thrombosis) and PE (Pulmonary Embolism)
are important for three reasons–managing the current event, identifying and
managing recurrent disease, and managing prophylaxis in high–risk situations such
as pregnancy or surgery. The initial objective tests are ultrasound for DVT and lung
scanning or helical CT for PE.
Primary heparin therapy is essential for VTE and until recently this was administered
by continuous intravenous infusion in hospital, followed by outpatient warfarin.
Low molecular weight heparins are now available which enable many patients with
both DVT and PE to be treated on an outpatient basis. The low molecular weight
heparin (LMWH) with dose adjusted for bodyweight, is given subcutaneously once
or twice each day for a minimum period of five days, followed by warfarin. Using this
approach, many patients can be managed as outpatients.
SeeThrombophilia
Capital Pathology Handbook – Interpretation of Laboratory Tests
Virology Swabs and Specimens
Please provide brief clinical details. If possible, state which viral infections you are
considering in the differential diagnosis.
Despite the vast spectrum of disease caused by viruses, they are less often
investigated by the laboratory than bacteria. They are harder to grow, harder to treat
and the trivial viral infections tend to be self–limiting.
Specific tests are described under the alphabetical entry for each disease.
Identification is by direct viral or virid antigen detection (PCR, immunofluorescence,
cell culture) or by detection of antibodies.
Virus or its antigen
Specimens can be faeces, throat swab, serum, vesicle fluid, CSF or aspirated
respiratory fluid. Swabs for viral investigation should be placed into viral transport
medium.
Antibodies
IgM antibodies usually indicate current infection. IgG antibodies indicate past or
present infection.
Diagnosis is established by either a single high titre or by an increase in titre of 4–fold
or more on paired sera, the first collected as early as possible in the disease, the
second after 2–3 weeks of illness.
Enquiries about specific tests can be directed to the Director of Clinical Pathology on
02 6285 9895.
Vitamin A (Retinol)
Specimen:Serum – Gel
Reference range: Supplied with report
V
A
Vitamin B12
Specimen:
Serum – Gel
Reference Range: > 170 pmol/L
Normal absorption of B12 requires a non–vegetarian dietary source, a normal stomach
to produce intrinsic factor, and a normal terminal ileum to absorb the B12/IF complex.
A healthy person with replete body stores has enough B12 to last 3–6 years if no more
is ingested.
Low B12 levels
Causes include:
•
•
•
•
•
•
•
•
•
•
•
•
Vegetarian diet
Drugs: – oral contraceptives
– metformin
– other: methotrexate, colchicine, Slow K, anticonvulsants,
cimetidine, triamterene
Pregnancy–B12 levels are often low without tissue deficiency
Low B12 binding protein, transcobalamin I, which can be measured, but the
expense is seldom warranted
Pernicious anaemia
Malabsorption
Inflammatory bowel disease: Crohn’s disease, or ulcerative colitis in the
terminal ileum
Previous gastrectomy or ileectomy
Primary folate deficiency
Diphyllobothrium fish tapeworm infestation.
Further investigation of low B12
If haematological abnormalities are present–raised MCV, anaemia, neutropenia,
thrombocytopenia, oval macrocytes and hypersegmented neutrophils in the blood
film–the underlying cause must be identified.
If there are no haematological abnormalities and the patient seems healthy, there
is less urgency. The level may be normal for that person or it may indicate early
deficiency. It is a matter of clinical judgement and patient choice whether to investigate
further at once or at follow–up. The elderly often have low B12 and folate levels and
may benefit from supplements.
Elevated B12 levels
• Oral or parenteral vitamin B12 supplement
• Haematological disorders: myeloproliferative disorders, leukaemias, high white
cell counts
• Liver disease.
See Pernicious Anaemia
Vitamin C
Specimen:Serum – Lithium heparin
Spin, separate and freeze serum.
Wrap in foil to protect from light.
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Vitamin D
Specimen:Serum – Gel
Patient should be fasting.
Reference Range:<50 Deficient
50–75 Intermediate
>75 Sufficient
Who is at risk of Vitamin D deficiency?
•
•
•
•
•
People with limited sun exposure
Dark skinned people especially if veiled
Malabsorption
Pregnant women
Patients on certain medication such as anticonvulsants or corticosteroids.
Vitamin E
Specimen:Serum – Gel
Spin, separate and freeze serum.
Wrap in foil to protect specimen from light.
Reference Range: Supplied with report
V
A
Vitamin K
Vitamin K is a fat–soluble vitamin which is essential for hepatic formation of
functionally active prothrombin (Factor II) and coagulation Factors VII, IX and X.
Although vitamin K is found in the diet, particularly green vegetables, the more
important source is the normal bacterial flora of the bowel which synthesises the
vitamin. Body stores last only 1–2 weeks if absorption stops.
Deficiencies of vitamin K and Factors II, VII, IX and X can be due to pre–hepatic or
hepatic causes.
The prothrombin ratio (PR) and its standardised presentation, INR, are used to
monitor vitamin K–dependent factor deficiencies and the anticoagulant effect of
warfarin.
Vitamin K injections correct factor deficiencies due to lack of absorption or availability
and are also used to correct a prolonged INR due to excess warfarin action.
SeeINR
Von Willebrand’s Disease
vWD, discovered in 1926 and the commonest inherited bleeding disorder, is due to a
defect in the plasma, von Willebrand factor (vWF) causing a secondary abnormality or
platelet adhesion.
vWF, like fibrinogen and fibronectin, is an adhesive protein important in platelet
attachment to subendothelial surfaces and in platelet–platelet interactions at sites of
vessel injury. vWF also plays an important role in the stabilisation and protection of
coagulant FVIII:C in plasma.
The most commonly encountered type of vWD is transmitted as an autosomal
dominant trait. These patients usually have mild bleeding symptoms and mild to
moderate abnormalities in the relevant laboratory tests (type I heterozygous vWD).
Testing for vWD
FVIII:C The coagulant activity of the FVIII:C protein.
von Willebrand Factor (vWF)
Measured as the von Willebrand factor antigen (vWF:Ag). This large glycoprotein
normally stabilises Factor FVIIIC and is variably reduced in vWD. vWF is essential
for normal adhesion of platelets and therefore for a normal bleeding time.
von Willebrand factor activity (vWF:Activity)
This is a functional activity which measures the vWF’s ability to bind to the
Glycoprotein Ib platelet binding site responsible for normal adhesion.
Collagen binding assay (CBA)
The functional assay which measures the vWF’s ability to bind to collagen. A
discrepancy between the vWF antigen and the CBA suggests a variant form of vWD.
Ristocetin–induced platelet aggregation
The functional activity of vWF measured as the ability of plasma vWF to agglutinate
platelets in the presence of ristocetin. The assay is relatively sensitive and specific for
vWD.
In the initial laboratory evaluation of patients suspected of having vWD, the following
should be performed:
Capital Pathology Handbook – Interpretation of Laboratory Tests
•
•
•
•
•
•
•
•
Bleeding time
Platelet count
APTT
FVIII:C
vWF Activity
CBA
Blood group
Ristocetin platelet aggregation (not routinely performed in all patients).
One or more of these tests are usually abnormal in patients who have a vWD but
the results may vary in the same patient with repeat testing. It is essential that the
patient not be taking drugs which could affect the bleeding time. Most frequent offending
agents are aspirin or other NSAIDs. Many conditions such as pregnancy, hypo–or
hyperthyroidism, uraemia, recent exercise, infection diabetes can affect the FVIII:C activity
and vWF antigen levels. Individuals with blood group O have significantly reduced levels
of the vWF:Ag and vWF activity compared with blood groups A, B or AB.
V
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
W
Wound Swabs
A well collected wound swab can provide valuable information. It is important to
sample from any discharging area, and to avoid touching outside the wound perimeter
thus avoiding skin contaminants. In the results interpretation the clinical appearance of
any wound is important, as well as considering the Gram stain result. Simply growing
an organism/s from a wound does not necessarily imply isolation of a significant
pathogen.
W
A
Capital Pathology Handbook – Interpretation of Laboratory Tests
X
X
A
Xylene
Specimen:
Urine post shift
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Y
Y
A
Yersinia enterocolitica
Specimen:Serum – Gel
A pathogen which is being isolated with increasing frequency from cases of infectious
diarrhoea–currently about 10% of the total.
Yersinia spp. are frequently present in the intestinal tract of wild and domesticated
clinical healthy birds and animals. Processed meat ready for sale has only rarely
been shown to be contaminated with Yersinia. The source of most cases of Yersinia
infection unknown.
Some patients develop an abdominal pain syndrome which may last for weeks. The
most likely cause for this is intra–abdominal lymphadenopathy.
Y. enterocolitica is susceptible to cotrimoxazole, tetracycline and fluoroquinolone but
antibiotics do not shorten the diarrhoea which usually settles spontaneously in 3–10
days.
Infections tend to be sporadic rather than epidemic and are found worldwide. It
can cause terminal ileitis which mimics acute appendicitis presenting as right lower
quadrant pain, fever and leucocytosis. Some patients develop a reactive arthritis.
Yersinia has been transmitted by blood donated by asymptomatic donors. Patients
who have Yersinia–associated diarrhoea should inform the blood collection service if
they donate blood over the next six months.
Yellow fever Antibodies
Specimen:
Serum – Gel
Reference Range: Supplied with report
Capital Pathology Handbook – Interpretation of Laboratory Tests
Z
Z
A
Zarontin (Ethosuximide)
Specimen: Plasma – Lithium heparin
Trough level is taken before next oral dose (within one hour).
Peak level is collected 2–4 hours after last oral dose.
Please supply time of last dose.
Reference Range: Supplied with report
Zinc, red cell
Specimen:
Whole blood – Trace element tube
Reference Range: Supplied with report
Zinc, serum
Specimen:
Serum – Gel
Reference Range: Supplied with report
Zinc, urine
Specimen:24–hour urine (nil preservative)
or
Spot urine–10 mL early morning.
Reference Range: Supplied with report
Capital Pathology, 2 Makin Place Deakin, ACT 2600 Australia
ABN 49 452 500 422
A subsidiary of Sonic Healthcare Limited
ABN 24 004 196 909
Contact us:
Doctors Service Centre (DSC) 02 6285 9803
www.capitalpath.com.au

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