CLS 444 Microbiology I

Antimicrobial Susceptibility Testing

Objectives: Upon completion of this CLS 444 lecture and lab, the Clinical Laboratory Science
student will:
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State the mode of activity, give examples, and describe the spectrum of activity for the major
antibiotic classes including:
a. Cell wall inhibitors (beta lactams and glycopeptides)
b. Alteration of cell membrane
c. Inhibitors of protein synthesis
d. Folic acid inhibitors
e. Inhibitors of nucleic acid synthesis
Differentiate between a bacteriostatic antibiotic and a bacteriocidal antibiotic.
Explain the purpose of antimicrobial therapy.
List factors that a clinician would take into consideration before prescribing an
antimicrobial agent for a patient.
Discuss the reasons tetracycline and the quinolones/fluoroquinolones are contraindicated in
children and pregnant women
Discuss the reason nitrofurantoin and norfloxacin is useful in treating urinary tract infections.
Discuss why metronidazole is useful in treating anaerobic infections.
-Activates under anaerobic conditions
Differentiate between narrow and broad spectrum antimicrobial agents.
Differentiate intrinsic and acquired resistance.
a. Discuss the means of acquisition of resistance mechanisms
b. Provide examples of each
Determine the role of the Clinical and Laboratory Standards Institute (CLSI) in the clinical
microbiology laboratory.
List four factors to consider if antimicrobial susceptibility testing is indicated.
-Is organism probably cause of infection
-Body site retrieved from
-Quality of specimen (pure or contaminants)
-Host status
Describe how drug formularies are chosen for health care institutions.
List the methods for susceptibility testing.
Explain the terms: susceptible, intermediate, and resistant.
Describe the principle, inoculation, required medium, incubation, reading and interpretation
of each of the following:
a. Kirby-Bauer (Disk diffusion)
b. E-Test
c. D test
d. ESBL screen
e. Oxacillin screen for penicillin resistance for Streptococcus pneumoniae
f. Aminoglycoside high level resistance for Enterococcus sp.
Define the following:
a. Minimum inhibitory concentration (MIC).
b. Minimum bactericidal concentration (MBC).
c. Zone of inhibition
d. Empirical therapy

16. Read and interpret the results (including the proper reporting) of disk diffusion, E-test, MIC, MBC,
ESBL screen, MRSA screen, VRE screen, and D test.
17. Differentiate between organisms that are susceptible, intermediate, or resistant to a particular
antimicrobial agent.

18. Describe how susceptibility testing is standardized and predict the outcome if a standardized
factor is altered for disk diffusion testing.
a. Bacterial inoculum and purity
b. Growth medium
i. pH
ii. Cation concentration-Ca+2 and Mg+2
iii. Agar depth
c. Incubation temperature, atmosphere, and duration
19. Define the following and state the clinical significance:
a. MRSA
b. VRE
c. VRSA
d. ESBL
e. Carbapenemases (KPC)
f. AmpC
20. Differentiate between synergism, antagonism, and indifference.
21. Define: serum bactericidal test (Schlichter test), peak level, and trough level.
22. Define beta-lactamase and explain the principle of the most used method to detect it (cefinase or
nitrocefin).
23. Define antibiogram
24. Recognize types of automated sensitivity testing systems.
25. Cite advantages of automated sensitivity testing systems.