Introduction to
DIAGNOSTIC
MICROBIOLOGY
for the Laboratory Sciences
Maria Dannessa Delost, PhD, MT(ASC-P)
Professor and Director of Medical Laboratory Programs
Department of Health Professions
The Bitonte College of Health & Human Services
Youngstown State University
Youngstown, OH
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Library of Congress Cataloging-in-Publication Data
Delost, Maria Dannessa, author.
Introduction to diagnostic microbiology for the laboratory sciences / Maria Dannessa Delost.
p. ; cm.
Includes bibliographical references and index.
ISBN 978-1-284-03231-4 (pbk.)
I. Title.
[DNLM: 1. Clinical Laboratory Techniques—Problems and Exercises. 2. Microbiological Techniques—Problems and
Exercises. QY 18.2]
RB38.25
616.07'5076—dc23
2013034746
6048
Printed in the United States of America
18 17 16 15 14 10 9 8 7 6 5 4 3 2 1
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Dedication
To the memory of my Dads, Amil M. Dannessa
and George Delost, Jr., who served their country
in World War II as members of our Greatest
Generation and who taught me the values of
hard work, loyalty, and honesty
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Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xv
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xvii
Reviewers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xix
Chapter 1
Introduction to Clinical Microbiology . . . . . . . . . . . . . . . . . . . . 1
Chapter 2
Safety in the Clinical Microbiology Laboratory . . . . . . . . . . . . . . 17
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Classification and Taxonomy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Characteristics of Eukaryotes and Prokaryotes . . . . . . . . . . . . . . . . . . . . . . . 3
Bacterial Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Cellular Metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Exotoxins and Endotoxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
The Role of Clinical Microbiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
The Infectious Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Normal Flora . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Modes of Transmission—Routes of Infection . . . . . . . . . . . . . . . . . . . . .8
True versus Opportunistic Pathogens . . . . . . . . . . . . . . . . . . . . . . . . . 9
Host Defense Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Signs of Infection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Nosocomial and Health Care–Associated Infections . . . . . . . . . . . . . . . . 14
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Laboratory Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Exposure Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Safety Education . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Universal Precautions and Standard Precautions . . . . . . . . . . . . . . . . . 19
Personal Protective Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Engineering Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Postexposure Plan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Fire Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Chemical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
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Contents
Chapter 3
Specimen Collection, Transport, and Processing: Preliminary
Identification Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
General Specimen Guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Specimen Collection Requirements for Specific Sites . . . . . . . . . . . . . . . . . . 33
Respiratory Tract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Gastrointestinal Tract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Urine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Blood . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Cerebrospinal Fluid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Genital Tract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Wound and Abscesses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Specimen Transport and Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Initial Specimen Plating and Identification Methods . . . . . . . . . . . . . . . . . . 38
Microscopic Examination of Clinical Specimens . . . . . . . . . . . . . . . . . . 38
Selection and Inoculation of Primary Media . . . . . . . . . . . . . . . . . . . . 39
Preliminary Biochemical Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Carbohydrate Utilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Catalase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Cytochrome Oxidase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Coagulase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Spot Indole . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
PYR Hydrolysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Chapter 4
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Microscopy, Staining, and Traditional Methods of Examination . . 51
Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Brightfield Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Direct Examination Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Staining Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Methylene Blue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Gram Stain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Acid-Fast Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Fluorescent Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Fungal Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Antibody-Conjugated Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
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Contents
Chapter 5
Automation, Immunodiagnostics, and Molecular Methods . . . . . 59
Multitest Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Automated Identification Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Principles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Automated Systems for Identification . . . . . . . . . . . . . . . . . . . . . . . . 62
Immunochemical Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Antigens and Antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Immunochemical Methods to Detect Microorganisms . . . . . . . . . . . . . . 65
Immunofluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Immunoserological Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Acute and Convalescent Phase Specimens . . . . . . . . . . . . . . . . . . . . . . 70
Immunoserological Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Introduction to Molecular Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Nucleic Acid Probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Amplification Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Nucleic Acid Sequencing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Chapter 6
Antimicrobial Susceptibility Testing . . . . . . . . . . . . . . . . . . . . 83
Chapter 7
Staphylococci and Other Catalase-Positive Gram-Positive Cocci . . 131
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Overview of Antibiotic Susceptibility Testing . . . . . . . . . . . . . . . . . . . . . . . 84
Selection of Antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Categories and Types of Antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Antimycobacterial Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Antifungal Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Antiviral Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Resistance to Antimicrobial Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Intrinsic Resistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Acquired Resistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Laboratory Methods for Susceptibility Testing . . . . . . . . . . . . . . . . . . . . . . 97
Disk Diffusion Susceptibility Testing . . . . . . . . . . . . . . . . . . . . . . . . . 98
Quantitative Dilution Susceptibility Testing . . . . . . . . . . . . . . . . . . . 100
Specific Methods to Determine Bacterial Resistance . . . . . . . . . . . . . . . 115
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Gram-Positive Cocci . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
Staphylococcus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Staphylococcus aureus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
Specimen Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
Infectious Processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
Detection of Antibiotic-Resistant Staphylococcus aureus . . . . . . . . . . . . 139
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Coagulase-Negative Staphylococci . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
Staphylococcus epidermidis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
Staphylococcus saprophyticus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148
Chapter 8
Streptococcus, Enterococcus, and Related Organisms . . . . . . . . . 149
Chapter 9
Neisseria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
Chapter 10
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Streptococcus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
Classification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
β-Hemolytic Streptococcus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
Streptococcus pyogenes (Group A Streptococcus) . . . . . . . . . . . . . . . . . . 152
Streptococcus agalactiae (Group B Streptococcus) . . . . . . . . . . . . . . . . . 155
Miscellaneous β-Hemolytic Streptococcus . . . . . . . . . . . . . . . . . . . . . . 156
Viridans Streptococci . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Streptococcus pneumoniae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
Streptococcus bovis Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
Enterococcus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
Nutritionally Variant Streptococci (NVS): Abiotrophia
and Granulicatella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Other Catalase-Negative Streptococcal-Like Organisms . . . . . . . . . . . . . 161
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
Characteristics of Neisseria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
Identification of Neisseria Species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
Infectious Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174
Neisseria gonorrhoeae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174
Neisseria meningitidis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178
Moraxella catarrhalis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
Miscellaneous Neisseria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
Enterobacteriaceae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Characteristics of Enterobacteriaceae . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
Serological Characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
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Identification and Significance of Selected Enterobacteriaceae . . . . . . . . . . . 194
Escherichia coli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
Shigella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
Klebsiella-Enterobacter-Serratia-Hafnia Group (Tribe Klebsielleae) . . . . . . 198
Proteeae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
Antibiotic Susceptibility and Treatment . . . . . . . . . . . . . . . . . . . . . . . . . 209
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223
Chapter 11
Nonfermentative Gram-Negative Bacilli . . . . . . . . . . . . . . . . 227
Chapter 12
Miscellaneous Gram-Negative Bacilli: Part 1 . . . . . . . . . . . . . . 245
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Characteristics of Nonfermenters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228
Identification of Nonfermenters . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
Family Pseudomonadaceae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231
Pseudomonas aeruginosa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231
Pseudomonas fluorescens and P. putida . . . . . . . . . . . . . . . . . . . . . . . 233
P. stutzeri and P. mendocina . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 234
Burkholderia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 234
Ralstonia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235
Acinetobacter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236
Stenotrophomonas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236
Alcaligenes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237
Achromobacter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237
Moraxella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238
Oligella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238
Chryseobacterium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238
Kingella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 239
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 239
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 242
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 242
Vibrio . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246
Vibrio cholerae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246
Other Vibrio . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247
Campylobacter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 248
Helicobacter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 250
Aeromonas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 250
Plesiomonas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252
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Chapter 13
Miscellaneous Gram-Negative Bacilli: Part 2 . . . . . . . . . . . . . . 255
Chapter 14
Gram-Positive Bacilli . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275
Haemophilus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256
Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256
Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 257
Haemophilus influenzae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 258
Miscellaneous Haemophilus Species . . . . . . . . . . . . . . . . . . . . . . . . 260
Legionella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 260
Bordetella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 262
Brucella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264
Pasteurella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265
Francisella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 266
Streptobacillus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267
HACEK Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267
Chromobacterium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 268
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 268
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 270
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 271
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 272
Spore Formers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
Bacillus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
Nonspore Formers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
Catalase Positive and Nonbranching . . . . . . . . . . . . . . . . . . . . . . . . 278
Catalase Negative and Simple Branching . . . . . . . . . . . . . . . . . . . . . . 281
Branching or Partially Acid Fast Gram-Positive Bacilli . . . . . . . . . . . . . 282
Miscellaneous Gram-Positive Bacilli . . . . . . . . . . . . . . . . . . . . . . . . . 283
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 284
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
Chapter 15
Spirochetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 287
Chapter 16
Anaerobes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
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Borrelia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 288
Relapsing Fever . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 288
Borrelia burgdorferi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289
Treponema . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291
Treponema pallidum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
Other Treponemes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 293
Miscellaneous Spirochetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 293
Leptospira . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 294
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
Overview of Anaerobes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300
Oxygen Tolerance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301
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Specimen Collection and Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301
Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
Anaerobic Culture Media and Incubation . . . . . . . . . . . . . . . . . . . . . . . . 303
Culture Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
Incubation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
Biochemical and Other Identification Methods . . . . . . . . . . . . . . . . . 305
Clinically Relevant Anaerobes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
Anaerobic Gram-Negative Bacilli . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
Anaerobic Gram-Positive Spore-Forming Bacilli: Clostridium . . . . . . . . . . . . . 310
Clostridium tetani . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 310
Clostridium botulinum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 310
Clostridium perfringens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311
Clostridium difficile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 312
Miscellaneous Clostridia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313
Anaerobic Gram-Positive Non–Spore-Forming Bacilli . . . . . . . . . . . . . . . . . . 313
Anaerobic Gram-Positive Cocci . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 314
Anaerobic Gram-Negative Cocci . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315
Antimicrobial Susceptibility Testing for Anaerobes . . . . . . . . . . . . . . . . . . . 316
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 317
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 319
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 320
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 322
Chapter 17
Mycobacterium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 325
Chapter 18
Chlamydia, Mycoplasma, and Rickettsia . . . . . . . . . . . . . . . . . . . . . 351
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Classification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 326
Specimen Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 327
Specimen Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 327
Staining . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 328
Culture Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 329
Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 329
Mycobacterium tuberculosis Complex . . . . . . . . . . . . . . . . . . . . . . . . . . . 332
Mycobacterium tuberculosis and Tuberculosis . . . . . . . . . . . . . . . . . . . 332
Photochromogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 337
Scotochromogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 338
Nonphotochromogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339
Rapid Growers: Mycobacterium Fortuitum Chelonei Complex . . . . . . . . 340
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 340
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 345
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 346
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 348
Chlamydiaceae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
Chlamydia trachomatis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
Chlamydophila psittaci . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 354
Chlamydophila pneumoniae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 354
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Mycoplasma . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 354
Rickettsiaceae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 356
Rickettsia rickettsii . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
Rickettsia akari . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
Rickettsia prowazekii . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
Rickettsia typhi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
Orientia tsutsugamushi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
Anaplasmataceae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
Ehrlichia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
Coxiella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360
Chapter 19
Introduction to Virology . . . . . . . . . . . . . . . . . . . . . . . . . . 363
Chapter 20
Introduction to Medical Mycology . . . . . . . . . . . . . . . . . . . . 395
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Viral Structure and Characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364
Viral Replication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365
Specimen Selection, Collection, and Processing . . . . . . . . . . . . . . . . . 367
Laboratory Methods in Virology . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
Serological Detection of Viral Antibodies . . . . . . . . . . . . . . . . . . . . . . 371
Selected Medically Important Viruses . . . . . . . . . . . . . . . . . . . . . . . . . . . 372
Adenovirus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 372
Hepatitis Viruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 372
Herpes Viruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 375
Papillomaviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 378
Papovaviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 379
Poxviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 380
Arboviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 380
Filoviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 383
Picornaviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 383
Orthomyxoviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 384
Paramyxoviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 385
Rhabdoviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 387
Reoviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 387
Caliciviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 387
Retroviridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 388
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391
Introduction to Mycology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 396
Specimen Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398
Identification Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398
Culture Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 402
Microscopic Examination of Growth . . . . . . . . . . . . . . . . . . . . . . . . 403
Incubation and Macroscopic Morphology . . . . . . . . . . . . . . . . . . . . . 403
Fungal Infections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 405
Superficial Mycoses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 405
Subcutaneous Mycoses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 412
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Systemic Mycoses: Dimorphic Fungi . . . . . . . . . . . . . . . . . . . . . . . . . 414
Yeast and Yeast-Like Fungi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 418
Candida . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 420
Opportunistic Saprobic Fungi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 423
Serological Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 425
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 426
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 429
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 431
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 432
Chapter 21
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Introduction to Medical Parasitology . . . . . . . . . . . . . . . . . . 435
Introduction and Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 437
Specimen Collecting, Processing, and Examining of Stool Specimens . . . . . . . 438
Collection and Processing of Stool Specimens . . . . . . . . . . . . . . . . . . 438
Microscopic Examination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439
Direct Smears . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439
Concentration Methods for Stool Specimens . . . . . . . . . . . . . . . . . . . 439
Permanently Stained Mounts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 440
Blood Specimens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 440
Protozoa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 440
Intestinal Amoebas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 441
Free-Living Amoebas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 441
Intestinal Flagellates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445
Intestinal Ciliate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 446
Intestinal Coccidia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 446
Miscellaneous Protozoa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 449
Sporozoa from Blood and Tissue . . . . . . . . . . . . . . . . . . . . . . . . . . . 450
Hemoflagellates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 456
Trypanosomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 456
Nematodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 458
Strongyloides stercoralis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 459
Hookworms: Necator americanus and Ancylostoma duodenale . . . . . . . 459
Enterobius vermicularis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 462
Ascaris lumbricoides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 463
Trichuris trichiura . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 465
Trichinella spiralis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 465
Blood and Tissue Nematodes: Filarial Worms . . . . . . . . . . . . . . . . . . . 468
Cestodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 470
Diphyllobothrium latum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 471
Hymenolepsis nana . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 473
Hymenolepsis diminuta . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 473
Dipylidium canium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 474
Taenia saginata, T. solium, and T. asiatica . . . . . . . . . . . . . . . . . . . . . 476
Echinococcus granulosus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 476
Trematodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 478
Intestinal Flukes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 479
Schisotosomes: Blood Flukes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 483
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Contents
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 485
Laboratory Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 489
Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 490
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 493
Chapter 22
Clinical Specimens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 495
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 496
Primary Plating of Specimens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 496
Blood Cultures and Bacteremia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 496
Types and Causes of Bacteremia . . . . . . . . . . . . . . . . . . . . . . . . . . . 497
Blood Culture Specimen Collection . . . . . . . . . . . . . . . . . . . . . . . . . 497
Manual Blood Cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 501
Automated Blood Culture Systems . . . . . . . . . . . . . . . . . . . . . . . . . 503
Microorganisms Associated with Blood Infections . . . . . . . . . . . . . . . . 503
Cerebrospinal Fluid—Meningitis and Central Nervous System Infections . . . . 504
Collection and Analysis of CSF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 507
Respiratory Tract Specimens and Infections . . . . . . . . . . . . . . . . . . . . . . . 509
Upper Respiratory Tract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 509
Throat Specimens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 509
Lower Respiratory Tract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 510
Diagnosis of Lower Respiratory Tract Infections—Sputum Specimens . . . 512
Eye and Ear Specimens and Infections . . . . . . . . . . . . . . . . . . . . . . . . . . . 514
Urinary Tract Specimens and Infections . . . . . . . . . . . . . . . . . . . . . . . . . . 514
Urine Collection and Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . 516
Colony Count . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 517
Automated Urine Screening Methods . . . . . . . . . . . . . . . . . . . . . . . . 518
Gastrointestinal Tract Specimens and Infections . . . . . . . . . . . . . . . . . . . . 518
Specimen Collection and Processing . . . . . . . . . . . . . . . . . . . . . . . . 520
Genital Tract Specimens and Infections . . . . . . . . . . . . . . . . . . . . . . . . . . 522
Specimen Collection for Urogenital Infections . . . . . . . . . . . . . . . . . . . 523
Direct Examination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 523
Skin and Wound Specimens and Infections . . . . . . . . . . . . . . . . . . . . . . . . 525
Specimen Collection for Wound and Skin Infections . . . . . . . . . . . . . . 526
Pleural, Peritoneal, Pericardial, and Synovial Fluids . . . . . . . . . . . . . . . . . . 526
Pleural Fluid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 528
Peritoneal Fluid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 528
Pericardial Fluid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 528
Synovial Fluid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 528
Laboratory Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 529
Case Studies and Review Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . 538
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 542
Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 545
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 559
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Preface
Introduction to Diagnostic Microbiology for the Laboratory Sciences provides a concise study of clinically significant microorganisms for the medical laboratory science
student and the laboratory professional. This text is on
the recommended reading list to prepare for the ASCP
MLT (American Society for Clinical Pathology, Medical
Laboratory Technician) exam. Although comprehensive
textbooks for diagnostic microbiology abound, this text
is unique in its focus on what is most essential for the beginning student. The concept of this text was originally
suggested by my medical laboratory technology students
who were frustrated by the length and depth of other
microbiology textbooks. Students became discouraged
with the vast amount of information included in these
texts, which serve as excellent learning resources but
were inappropriate for the beginning student.
This text provides an overview of diagnostic microbiology that is relevant and essential to medical
laboratory students and entry level medical laboratory
technicians. Chapters 1 through 6 begin by providing
an overview of the infectious process, including disease transmission and host immunity. The importance
of specimen collection is emphasized with both general and specific requirements, instilling in the student
the importance of pre-analytical factors in specimen
management and quality laboratory testing. The goal
is to provide a thorough yet succinct discussion of isolation media, as well as the basics of preliminary testing in the microbiology laboratory.
Essential manual methods are addressed both in
theory and method so the student can understand,
practice, and apply each laboratory procedure. The
chapters mesh didactic theory with laboratory testing and interpretation, creating an orderly and complete presentation. Additional testing methods, such
as serology, immunoassay, antibiotic susceptibility
testing, and molecular diagnostics are also presented
with pertinent applications. Current modes of antibiotic resistance and the required testing are included
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which introduce the student to the increased attention
required in antibiotic testing, as well as the identification of resistance patterns. The expanded information
on molecular diagnostics provides the student with
the essential framework and examples of the expanding use of molecular technologies in the microbiology laboratory. The text also introduces the student
to current automation in the microbiology laboratory,
including contemporary instrumentation used in microbial identification, antibiotic susceptibility testing,
and blood culture technologies.
Chapters 7 through 18 address medically important bacteria, including their isolation, identification, and treatment. Each chapter focuses on major
divisions of bacteria, including gram-positive cocci,
gram-negative cocci, gram-negative bacilli, grampositive bacilli, mycobacterium, anaerobes, spirochetes, chlamydia, and rickettsia. Each chapter includes
concise information on the isolation, identification,
and clinical relevance of those microorganisms that a
microbiologist may encounter in the clinical setting.
These chapters also include news pieces that describe
a timely concern related to a specific microorganism
and its associated public health concern. Examples
include antibiotic resistance in Staphylococcus aureus,
meningococcoal disease, listeriosis, Lyme Disease, and
legionellosis. These items attempt to instill in the student the relevance of clinical microbiology in their
everyday life by relating contemporary events to the
lessons presented in the chapter.
Chapters 19 through 21 provide an orderly presentation of clinical virology, mycology, and parasitology
that can be realistically covered in an introductory
course. These topics are often overwhelming in both
quantity and depth of information in most microbiology texts, leading to frustration in many students. These important topics are summarized in
a format that encourages the student to review and
apply central concepts in each discipline. Chapter 19,
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Preface
Introduction to Virology summarizes viral replication, classification of DNA and RNA viruses, and the
isolation and identification of medically important
viruses. This chapter also summarizes and describes
significant viral infections and the role of the laboratory in the diagnosis of viral infections. Chapter 20,
Introduction to Medical Mycology discusses the classification of mycoses, specimen collection, and fungal
culture media. Clear identification schemes utilizing
both macroscopic colonial appearance and microscopic characteristics as well as biochemical methods
for medically important fungi are included. Superficial, subcutaneous, systemic, and opportunistic
mycoses are addressed. This chapter includes images that are vital in the correct identification of the
fungi. Chapter 21, Introduction to Medical Parasitology provides an organized classification outline
of medically relevant parasites and infections. Specimen collection requirements unique to parasitology
are described as well as concentration methods and
stains. The chapter is divided into units which discuss
protozoans such as the amebae, sporozoans and hemoflagellates, nematodes, cestodes, and trematodes.
Extensive, clearly drawn life cycles with images of the
infectious stage and diagnostic stage are included for
clinically relevant parasites.
The final chapter, Chapter 22, Clinical Specimens, ties the components of the text together by
summarizing the medically important pathogens typically found in clinical specimens with identification
schemes. Complete work-ups of microorganisms from
major clinical sites are included with flowcharts and
suggested laboratory protocols. Students are able to
apply the didactic and laboratory concepts presented
throughout the text to evaluate clinical specimens and
develop appropriate testing schemes to correctly identify pathogens and differentiate these from normal
flora.
The goal of the text is to provide the medical laboratory student with the information and skills necessary for entry into the medical laboratory profession.
The information is organized into a format that facilitates learning through the use of integrated text,
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charts, figures, and laboratory exercises. This format
also provides a quick reference to the most frequently
utilized manual procedures and most commonly
referred topics in clinical microbiology.
Key Features
Introduction to Diagnostic Microbiology for the Laboratory Sciences includes learning objectives, laboratory
exercises, news features, and review questions. Both
didactic and psychomotor skills are presented and reinforced in each chapter.
77 Learning Objectives–Clearly outlines for the reader
the desired outcomes of the chapter, focuses reading, and provides a basis to gauge learning.
77 Laboratory Exercises–Engages the reader by having them apply what they learned in a lab activity,
record their results, and push their analysis further with critical thinking questions.
77 News Feature–Highlights current issues that face
the contemporary medical laboratory technician,
grounding the information in a real-word context.
77 Review Questions–Provides a way to apply what
has been learned in the chapter and also provides
practice for preparation for the Medical Lab Technician (MLT) certification examination.
Many reference texts in clinical microbiology are
available with complex descriptions and highly detailed methods which can intimidate the beginning
student. The purpose of Introduction to Diagnostic
Microbiology for the Laboratory Sciences is to provide
a condensed and practical approach for students and
novice microbiologists. A mastery of the material and
procedures in this text will provide students with a
strong background for pursuing higher levels in clinical microbiology. Additionally, the text is an excellent resource to prepare for the ASCP certification
examination. It further serves as a valuable review for
laboratory professionals who are returning to the microbiology field after an absence. The concise format
provides a quick and thorough review mechanism for
students and laboratory professionals who may need a
refresher course in diagnostic microbiology.
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Acknowledgments
Special thanks to my husband, Raymond, and my sons, Gregory and Michael, and my entire family for their
encouragement and support during this project. Also, thank you to all of my medical laboratory students at
Youngstown State University from whom I have learned so much during my thirty years of teaching.
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Reviewers
Angela R. Bell, MS, MLS(ASCP)SM, DLM
Tidewater Community College
Virginia Beach, VA
Janet E. Cooper, MSNS, BSMT(ASCP)H
Instructor in Medical Laboratory Technology
Mississippi Delta Community College
Moorhead, MS
Katrina P. Ghazanfar, PhD, MT(ASCP)
University of Hawaii-Kapiolani
Community College
Honolulu, HI
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Michelle M. Hill, MS, MT(ASCP)
Instructor, MLT Program
Southwest Tennessee Community College
Memphis, TN
Benjamin F. Newsome Jr., MA/MS, MT(AMT), CM
Director
San Jacinto College
Pasadena, TX
Dale Telgenhoff, PhD, HTL(ASCP)
Associate Professor
Tarleton State University
Fort Worth, TX
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