Basic laboratory procedures in clinical bacteriology

Basic laboratory procedures in clinical bacteriology

 Contents

Preface viii

Introduction 1

Quality assurance in bacteriology 2

Introduction 2

Definitions 2

Internal quality control 6

External quality assessment 16


PART I : Bacteriological investigations 19

Blood 20

Introduction 20

When and where bacteraemia may occur 20

Blood collection 20

Blood-culture media 22

Processing of blood cultures 23

Cerebrospinal fluid 25

Introduction 25

Collection and transportation of specimens 25

Macroscopic inspection 26

Microscopic examination 26

Preliminary identification 28

Susceptibility testing 29

Urine 30

Introduction 30

Specimen collection 30

Culture and interpretation 32

Interpretation of quantitative urine culture results 34

Identification 35

Susceptibility tests 36

Stool 37

Introduction 37

Etiological agents and clinical features 37

Appropriate use of laboratory resources 39

Collection and transport of stool specimens 40

Visual examination of stool specimens 41

Enrichment and inoculation of stool specimens 41

Media for enteric pathogens 42

Primary isolation 42

Preliminary identification of isolates 44

Final microbiological identification 50

Serological identification 54

Upper respiratory tract infections 60

Introduction 60

Normal flora of the pharynx 60

Bacterial agents of pharyngitis 61

Collection and dispatch of specimens 62

Direct microscopy 62

Culture and identification 63

Susceptibility testing 65

Lower respiratory tract infections 66

Introduction 66

The most common infections 66

Collection of sputum specimens 68

Processing of sputum in the laboratory (for

non-tuberculous infections) 68

Culture for Mycobacterium tuberculosis 72

Interpretation of cultures for M. tuberculosis 74

General note on safety 74

Sexually transmitted diseases 76

Introduction 76

Urethritis in men 77

Genital specimens from women 79

Specimens from genital ulcers 82

Purulent exudates, wounds and abscesses 86

Introduction 86

Commonly encountered clinical conditions and the

most frequent etiological agents 86

Collection and transportation of specimens 89

Macroscopic evaluation 90

Microscopic examination 91

Culture 92

Identification 93

Susceptibility testing 97

Anaerobic bacteriology 98

Introduction 98

Description of bacteria in relation to oxygen requirement 98 

Bacteriology 98

Antimicrobial susceptibility testing 103

Introduction 103

General principles of antimicrobial susceptibility testing 103

Clinical definition of terms “resistant” and “susceptible”:

the three category system 104

Indications for routine susceptibility tests 106

Choice of drugs for routine susceptibility tests in the

clinical laboratory 107

The modified Kirby–Bauer method 109

Direct versus indirect susceptibility tests 117

Technical factors influencing the size of the zone in the

disc-diffusion method 118

Quality control 120

Serological tests 122

Introduction 122

Quality control measures 122

Serological reactions 125

Serological tests for syphilis 126

Febrile agglutinins tests 133

Antistreptolysin O test 135

Bacterial antigen tests 137


PART II: Essential media and reagents 141

Introduction 142

Pathogens, media and diagnostic reagents 143

Blood 144

Cerebrospinal fluid 144

Urine 145

Stool 146

Upper respiratory tract 147

Lower respiratory tract 148

Urogenital specimens for exclusion of sexually transmitted

diseases 149

Pus and exudates 149

List of recommended media and diagnostic reagents for the intermediate microbiological laboratory 150

Selected further reading 154

Index 155