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Preface to the Third Edition
In the last 27 years, since the publication of the first edition of Medical Laboratory Technology, many
changes in the field of laboratory medicine have occurred. Although some of these updates were included
in our second edition (2010), we felt that the time has come for a thorough revision, and thus we bring forth
the current edition.
When I look back, at my age of ninety, I can see how far medical technology has come through these
years. I could never forget the limited facilities I had for teaching science in India early in my career. It
was only after I moved to the United States that I was exposed to advanced medical laboratory education.
After gaining years of teaching experience on the subject here, I felt compelled to share what I learned
with my people whom I had left behind. Endless support came from my new friends in the United States
as well as in India with whom I worked during my tenure as a Fulbright Professor to lecture on Medical
Laboratory Technology across India. The best support was from the readers and academic institutions of
developing countries, who honoured our honest efforts. This would have not been possible without the help
from numerous doctors from rural and urban regions of India and people who were associated with the most
advanced laboratories in the United States. Thus the book stands as a model of collaboration in the field of
medical technology between those who truly understand the needs of the developing world and those who
have the knowledge of the developed world. I acknowledge with a deep sense of gratitude, my friends,
colleagues, students and members of my own family who have contributed to this bold endeavour.
As I worked with two dozen young authors in this edition, I could feel their motivation and eagerness to
share their hard-earned experience with new students, no matter how far away they may be in the world. As
they worked with me, they inspired me at every step and I felt young again. We, as a team, kept in mind that
the economic condition of developing countries will not permit the clinical laboratories to run on par with
advanced laboratories. Yet, the needs of the sick must be addressed and a doctor needs accurate laboratory
information to make the correct diagnosis. Hence, the laboratories of developing countries, including
those located far away from urban centres, have to operate on a limited budget and depend on resourceful,
knowledgeable laboratory technologists, who may have limited training or education.
Unfortunately, as I have observed, there remains a significant lack of professionalism in the clinical
laboratories of many developing countries. This has resulted in unreliable laboratory reports that can then
lead to wrong diagnoses, and as a result, unnecessary illness and death. A firm understanding of the critical
role of laboratories in alleviating human suffering may help increase the care and diligence with which
laboratory technicians perform their jobs. I hope and pray that this book can help make that change.
Kanai L Mukherjее
Professor Emeritus
Fulbright Visiting Professor to India (1983-1984)
Essex Community College,
Contents
Foreword vii
Preface to the Fourth Edition ix
Preface to the Third Edition x
Contributors xvii
Commonly Used Abbreviations in Medical Laboratories xix
SECTION 4: MICROBIOLOGY AND VIROLOGY
19. Introduction to Diagnostic Microbiology 471
Kanai L Mukherjee, Monisha Bhatia and Ritwik Bhatia
Part 1: Introduction to Infectious Diseases 472
Part 2: Diagnostic Microbiology 473
Role of Microbiology Laboratory 473
Rules of Microbiology Laboratory 473
Basic Laboratory Procedures in Microbiology 482
Microscopic Examination 485
Examination of Stained Smear (Dry Mount) 489
Laboratory Culture 500
Inoculation in Test Tubes 516
Culture Environments 517
Quality Control in Microbiology 521
Preservation of Stock Cultures 521
Identification of Pathogenic Microbes 523
Quick Reference of Media and Biochemical Tests 527
Part 3: Virology 530
Introduction 530
Diseases Caused by Viruses 530
Defence Mechanism of Body Against Viral Infection 531
Part 4: Laboratory Diagnosis of Viral Infections 531
Direct Examination 532
Serological Testing 532
Limitations of Serological Diagnosis 542
Review Questions 542
20. Identification of Pathogenic Bacteria 544
Ritwik Bhatia and Shovan Bhatia
Introduction to Diagnostic Bacteriology 545
Systematic Grouping of Pathogenic Bacteria 547
Specific Bacteria 549
Laboratory Identification of Infectious Agents 553
Laboratory Diagnosis of Leprosy 587
Diagnosis of Anaerobic Infections 589
Medical Laboratory Technology: Volume 2
xii
Identifying Characteristics of Common Pathogenic Bacteria 591
Antimicrobial Susceptibility Testing 606
Review Questions 614
Appendix 20.1: Formulae and Preparation of Culture Media and Biochemical Reagents 614
21. Laboratory Diagnosis of Mycotic Infections 644
Monisha Bhatia and Kanai L Mukherjee
Introduction to Fungi and Mycotic Agents 644
Clinical Significance of Mycotic Infection 645
Morphological Features of Mycotic Agents 646
Opportunistic Fungi and Filamentous Bacteria 648
Laboratory Diagnosis of Mycotic Infections 649
Mycotic Agents Classified by Location 662
Traveller's Mycoses 663
Identifying Characteristics of Common Pathogenic Fungi 663
Microbiology Laboratory Update 675
Review Questions 677
22. Laboratory Diagnosis of Parasitic Infections 678
Kanai L Mukherjee and Piyali Basu
Introduction to Parasites 679
Essential Background 680
Laboratory Procedures in Parasitic Infection 682
Laboratory Diagnosis of Pinworm Infections 703
Laboratory Investigation of Other Specimens for Parasitic Infections 705
Laboratory Diagnosis of Protozoan Infections 708
Laboratory Diagnosis of Parasitic Infections 712
Laboratory Diagnosis of Helminth Infections 716
Identification Characters of Helminths 717
Review Questions 724
Morphology and Life Cycles of Human Parasites i
SECTION 5: SEROLOGY
23. Introduction to Immunology and Principles of Serodiagnosis 729
Ashoke Khanwalkar and Raktima Datta
The Human Immune System 729
Immunology and Serology 735
Blood Types and Compatibility Testing 736
Serodiagnosis 736
Principles of Serological Procedures 737
Visible Immunologic Reactions Used in Serodiagnosis 739
Problems in Serological Assays 747
Review Questions 749
24. Laboratory Procedures in Serology 750
Kanai L Mukherjee and Chhotelaal Pande
Quality Management in Serology Laboratory 751
Collection and Preparation of Specimen 752
Sexually Transmitted Infections and their Serology 754
Nontreponemal Test 756
Treponemal Test 763
Agglutination Tests of Serodiagnosis 768
Contents
xiii
Serodiagnosis of Streptococcal Infection 780
Additional Serodiagnosis Kits 784
Infectious Mononucleosis and Monospot Test 785
Serodiagnostic Tests for Miscellaneous Infections 786
Immunologic Test for Pregnancy 797
Review Questions 801
SECTION 6: CLINICAL PATHOLOGY
25. Urine Analysis 805
Bibhas Bandy
Laboratory Examination of Body Fluids 805
Clinical Significance of Urinalysis 806
Laboratory Diagnosis of Renal Function 810
Routine Examination of Urine 814
Review Questions 844
26. Laboratory Examination of Miscellaneous Body Fluids 845
Bibhas Bandy
Body Fluids for Clinical Diagnosis 846
Cerebrospinal Fluid (CSF) 847
Serous Fluids 861
Synovial Fluid 864
Vaginal Discharge 867
Gastric Juice 869
Review Questions 870
27. Semen Analysis 872
Bibhas Bandy
Introduction 872
Laboratory Investigation 875
Forensic Investigation for the Presence of Semen 880
Review Questions 881
28. Stool Examination 882
Bibhas Bandy
Introduction 882
Laboratory Investigation 884
Processing of Watery Stool Specimen during Cholera Epidemic 893
Laboratory Analysis 894
Review Questions 898
Molecular Pathology i
Index I.1
CONTENTS OF VOLUME 1
SECTION 1: INTRODUCTION
1. Human Health and Clinical Diagnosis in Developing Countries 3
2. Introduction to Clinical Laboratories 16
Medical Laboratory Technology: Volume 2
xiv
3. Laboratory Safety and First Aid 29
4. Introduction to Laboratory Equipment and Basic Laboratory Operations 54
5. Specimen Handling and Laboratory Records 140
6. Units of Measurement and Preparation of Reagent Solutions 196
7. Good Laboratory Practices and Statistical Quality Control 212
SECTION 2: HAEMATOLOGY AND COAGULATION
8. Introduction to Haematology 229
9. Basic Laboratory Procedures in Haematology 236
10. Routine Haematological Tests 242
11. Special Haematological Tests 306
12. Interpretation of Laboratory Findings in Haematology 332
13. Introduction to Haemostasis and Haemostatic Disorders 338
14. Laboratory Investigation of Bleeding Disorders 348
SECTION 3: IMMUNOHAEMATOLOGY OR BLOOD BANKING
15. Introduction to Blood Transfusion Therapy 373
16. Collection and Processing of Blood for Transfusion 393
17. Routine Laboratory Procedures in Blood Bank 420
18. Blood Transfusion Services and Clinical Approach to
Haemolytic Disease of the Newborn 459
Laboratory Information Systems i
Phlebotomy/Venipuncture Procedure ix
Index I.1
CONTENTS OF VOLUME 3
SECTION 7: CLINICAL BIOCHEMISTRY
29. Biochemical Processes of the Body Under Normal and Pathogenic Conditions 901
Vissagan Gopalakrishnan and Piyali Basu
30. Specimen Collection and Processing for Biochemical Analyses 920
Chhotelaal Pandey and Anant Kumar Pande
31. Techniques of Analytical Chemistry 932
Kanai L Mukherjee and lshita Bhattacharya
xv
xv
32. Automation in Clinical Biochemistry 961
Aurin Chakravarty, Dipto Chakravarty and Chhotelaal Pandey
33. Routine Biochemical Test Procedures 985
Kanai L Mukherjee, Chhotelaal Pande and Rohini Chakravarthy
34. Biochemical Test Profiles 1081
Ashoke Khanwalkar
35. Therapeutic Drug Monitoring and Clinical Toxicology 1102
Kanai L Mukherjee and Piyali Basu
SECTION 8: HISTOLOGY AND CYTOLOGY
36. Introduction to Histotechnology and Cytotechnology 1127
Venk Mani and Surajit Ghosh
37. Laboratory Techniques in Histology 1147
Venk Mani and Kanai L Mukherjee
38. Laboratory Techniques in Diagnostic Exfoliative Cytology 1206
Krishna Mallik and Kanai L Mukherjee
39. Basics of Immunohistochemistry 1222
40. Polymerase Chain Reaction in Clinical Diagnosis 1240
SECTION 9: MISCELLANEOUS INFORMATION
Medical Terminology 1251
Maya Chattoraj and Kanai L Mukherjee
Glossary of Technical Terms 1254
Maya Chattoraj and Kanai L Mukherjee
Appendices 1284
Maya Chattoraj and Kanai L Mukherjee
Index I.1–I.8
Contributors
Bibhas Bandy, MD
Otolaryngologist (ENT Specialist)
Washington County Hospital
Hagerstown, MD, USA
Piyali Basu
, MS
Research Associate
Kolkata University, India
Monisha C Bhatia
Vanderbilt University, USA
Ritwik Bhatia
Vanderbilt University, USA
Shovan Bhatia
Research Associate
Ransom Everglades School
Miami, Florida, USA
Ishita Bhattacharya, BS
Research Associate
Monmouth Medical Scholar
Monmouth University, New Jersey, USA
Rohini Chakravarthy
Research Associate
Vanderbilt University Medical Centre
Tennessee, USA
Anuradha Chakravarthy
, MD
Professor, Department of Radiation Oncology
Vanderbilt University Medical Centre
Tennessee, USA
Aloka Chakravarthy
, PhD
Head of Statistics Department
US Food and Drug Administration
Washington, DC, USA
Aurin Chakravarty
, BS
Research Associate
Johns Hopkins University
Baltimore, MD, USA
Dipto Chakravarty
, MS
Specialist in Computer Technology
Washington, DC, USA
Tara Chattoraj
, BS
Research Associate
University of Illinois
Urbana Champaign, USA
Maya Chattoraj
Research Associate
Wheaton Warrenville South High School
Illinois, USA
Volkmar Dierolf
, PhD
Professor and Head
Department of Physics
Lehigh University, Allentown,
Pennsylvania, USA
Swarajit Ghosh
, MD
New Delhi, India
Vissagan Gopalakrishnan
Rush Medical College
Chicago, Illinois, USA
Ashoke Khanwalkar
, MD
Northwestern University
Chicago, Illinois, USA
Krishna Maliik
, MD
Specialist in Orthopaedic Surgery and Arthroscopy
Reconstruction, Mountain Vista Medical Centre
Mesa, Arizona, USA
Venk Mani
, MD
Chief Pathologist
Dickson Medical Associates
Dickson, Tennessee, USA
Kanai L Mukherjee
, MT, PhD
Fulbright Professor (1983),
Emeritus Retired Director of MLT Program (ECC)
Baltimore, MD, USA
Chhotelaal Pande
(NCA)
, BSc, MLT (ASCP), CLSH (NCA), CLSpH
Formerly Clinical Laboratory Supervisor,
Dar es Salaam, Tanzania, and
Metropolis Clinical Pathology Laboratory
Mumbai, Maharashtra, India
Medical Laboratory Technology: Volume 2
xviii
Anant Kumar Pandey, BS (Medical Biotechnology)
Amity University, Noida, Uttar Pradesh, India
Jay Raval
, MD
Assistant Professor
Medical Director of Therapeutic Apheresis
Division of Transfusion Medicine
University of North Carolina School of Medicine
Pampee Paul Young
, MD, PhD
Professor and Director
Blood Bank
Stem Cell Laboratory and Transfusion Services
Vanderbilt University Medical Centre
Nashville, Tennessee, USA
Commonly Used Abbreviations in Medical Laboratories
Rohini Chakravarthy
Abbreviation Expanded Version
A
Ab
ACT
AFB
Ag
AHG
AIDS
A
bsorbance
A
ntibody
A
ctivated clotting time
A
cid-fast bacillus
A
ntigen
A
ntihuman globulin
A
cquired immunodeciency
syndrome
ALL
ALP, AP
ALT
AML
ANA
APTT
A
cute lymphocytic leukemia
A
lkaline phosphatase
A
lanine aminotransferase
(formerly, SGPT)
A
cute myelogenous leukaemia
A
nti-nuclear antibody
A
ctivated partial
thromboplastin
AST
A
spartate aminotransferase
(formerly, SGOT)
BA
BBP
BP
BSC
BSI
BT
BUN
С
CBC
cc
CGL
chol
CK
CLL
B
lood agar
B
lood borne pathogen
B
lood pressure
B
iological safety cabinet
B
ody substance isolation
B
leeding time
B
lood urea nitrogen
C
elsius, centigrade
C
omplete blood count
C
ubic centimetre
C
hronic granulocytic leukaemia
C
holesterol
C
reatine kinase
C
hronic lymphocytic leukaemia
Abbreviation Expanded Version
CLS
CLT
cm
CNS
CO
CO
CPD
CPK
crit
C&S
CSF
2
C
linical laboratory scientist
C
linical laboratory technician
C
entimetre
C
entral nervous system
C
arbon monoxide
Carbon dioxide
C
itrate-phosphate-dextrose
C
reatinine phosphokinase
H
aematocrit
C
ulture and sensitivity
C
erebrospinal uid, colony
stimulating factor
cu mm
DAT
DIC
C
ubic millimetre (mm3)
D
irect antiglobulin test
D
isseminated intravascular
coagulation
Di
EBV
EDTA
EIA
EMB
ESR
EU
F
FBS
FDP
D
ierential count of WBC
E
pstein-Barr virus
E
thylenediaminetetraacetic acid
E
nzyme immunoassay
E
nzyme immunoassay
E
rythrocyte sedimentation rate
E
hrlich units
F
arenheit
F
asting blood sugar
F
ibrinogen degradation
products
FUO
G
g
GGT
GI
F
ever of unknown origin
G
ravitational force in dynes/cm
G
ram
G
amma glutamyl transferase
G
astrointestinal