The authors have emphasized the purposes and function of the clinical microbiology laboratory in the diagnosis of infectious diseases. The exercises illustrate as simply as possible the nature of laboratory procedures used for isolation and identification of infectious agents as well as the principles of asepsis, disinfection and sterilization. Attention is given to the role of the health professional in regard to appropriate collection of clinical specimens and the applications of aseptic and disinfectant techniques as they relate to patient care.

1. The Microscope

2. Handling and Examining Cultures

3. Simple Stains, Hanging Drop, and Wet-Mount Preparations

Experiment 3.1 Preparing a Simple Stain

Experiment 3.2 Preparing a Hanging Drop

Experiment 3.3 Preparing a Wet Mount

4. Gram Stain

5. Acid-Fast Stain

6. Special Stains

Experiment 6.1 Staining Bacterial Endospores (Schaeffer-Fulton Method)

Experiment 6.2 Flagella and Capsule Stains

7. Culture Media

8. Streaking Technique to Obtain Pure Cultures

9. Pour Plate and Subculture Techniques

Experiment 9.1 Pour-Plate Technique

Experiment 9.2 Subculture Technique (Picking Isolated Colonies for Pure Cultures)

PART TWO Destruction of Microorganisms

Physical Antimicrobial Agents

Chemical Antimicrobial Agents

Exercises

10. Moist and Dry Heat

Experiment 10.1 Moist Heat

Experiment 10.2 Dry Heat

Experiment 10.3 Incineration

11. The Autoclave

12. Disinfectants

Case Study: A Lesson Learned—Follow Proper Procedures for the Disinfection of Equipment

13. Antimicrobial Agent Susceptibility Testing and Resistance

Experiment 13.1 Disk Agar Diffusion Method

Experiment 13.2 Broth Dilution Method: Determining Minimum Inhibitory Concentration (MIC)

Experiment 13.3 Bacterial Resistance to Antimicrobial Agents

PART THREE Diagnostic Microbiology in Action

General Considerations

Microbiology at the Bedside

Collection and Transport of Clinical Specimens for Culture

Precautions for Handling Specimens and Cultures

Normal Flora of the Body

Principles of Diagnostic Microbiology

Exercises

14. Primary Media for Isolation of Microorganisms

15. Some Metabolic Activities of Bacteria

Experiment 15.1 Simple Carbohydrate Fermentations

Experiment 15.2 Starch Hydrolysis

Experiment 15.3 The Indole Test

Experiment 15.4 Use of Kligler Iron Agar

16. Activities of Bacterial Enzymes

Experiment 16.1 The Activity of Urease

Experiment 16.2 The Activity of Catalase

Experiment 16.3 The Activity of Oxidase

Experiment 16.4 The Activity of Deoxyribonuclease (DNase)

Experiment 16.5 The Activity of a Deaminase

17. Principles of Antigen Detection and Nucleic Acid Assays for Detection and Identification of Microorganisms

18. Staphylococci

Experiment 18.1 Isolation and Identification of Staphylococci

Experiment 18.2 Staphylococci in the Normal Flora

Case Study: Community-Acquired Methicillin-Resistant Staphylococcus

19. Streptococci, Pneumococci, and Enterococci

Experiment 19.1 Identification of Group A Streptococci (Streptococcus pyogenes)

Case Study: A Case of Pharyngitis

Experiment 19.2 The CAMP Test for Group B Streptococci (Streptococcus agalactiae)

Experiment 19.3 Identification of Pneumococci (Streptococcus pneumoniae)

Case Study: A Case of Pneumonia with Meningitis

Experiment 19.4 Identification of Enterococci

Experiment 19.5 Streptococci in the Normal Flora

20. Haemophilus, Corynebacteria, and Bordetella

Experiment 20.1 Haemophilus

Experiment 20.2 Corynebacteria

Experiment 20.3 Bordetella

Case Study: A School-Associated Pertussis Outbreak

21. Clinical Specimens from the Respiratory Tract

Experiment 21.1 Laboratory Diagnosis of a Sore Throat

Experiment 21.2 Laboratory Diagnosis of Bacterial Pneumonia

Experiment 21.3 Antimicrobial Susceptibility Test of an Isolate from a Clinical Specimen

22. The Enterobacteriaceae (Enteric Bacilli) and Other Clinically Important Gram-Negative Bacilli

Experiment 22.1 Identification of Pure Cultures of Enterobacteriaceae from the Normal Intestinal Flora

Experiment 22.2 Isolation Techniques for Enteric Pathogens

Case Study: A Case of Diarrhea

Experiment 22.3 Identification Techniques for Enteric Pathogens

Experiment 22.4 Serological Identification of Enteric Organisms

Experiment 22.5 Techniques to Distinguish Nonfermentative Gram-Negative Bacilli from Enterobacteriaceae

Experiment 22.6 Rapid Methods for Bacterial Identification

23. Clinical Specimens from the Intestinal Tract

Experiment 23.1 Culturing a Fecal Sample

Experiment 23.2 Identification of an Unknown Enteric Organism

Experiment 23.3 Antimicrobial Susceptibility Test of an Enteric Organism

24. Urine Culture Techniques

Experiment 24.1 Examination and Qualitative Culture of Voided Urine

Experiment 24.2 Quantitative Urine Culture

Case Study: A Case of Cystitis

25. Neisseria and Spirochetes

Experiment 25.1 Neisseria

Experiment 25.2 Spirochetes

Case Study: A Case of a Patient with a Bull’s-Eye Skin Lesion

26. Clinical Specimens from Blood

Case Study: Three Patients with Positive Blood Cultures

PART FOUR Microbial Pathogens Requiring Special Laboratory Techniques; Serodiagnosis of Infectious Disease; Principles and Practices of Infection Prevention

Exercises

27. Anaerobic Bacteria

Case Study: A Case of Diarrhea in a Hospitalized Patient

28. Mycobacteria

Experiment 28.1 Microscopic Morphology of Mycobacterium tuberculosis

Experiment 28.2 Culturing a Sputum Specimen for Mycobacteria 254

Case Study: A Case of Pneumonia in a Patient with Chronic Alcoholism

29. Mycoplasmas, Rickettsiae, Chlamydiae, Viruses, and Prions

30. Fungi:Yeasts and Molds

31. Protozoa and Animal Parasites

32. Serodiagnosis of Infectious Disease

33. Principles and Practices of Infection Prevention

Experiment 33.1 Nasal Surveillance Cultures for MRSA