Antimicrobial Susceptibility and Testing

Chemotherapy
• Chemotherapy refers to the use of drugs to treat a disease
• Selective toxicity: A drug that kills harmful microbes without damaging the
host
• Antibiotic: Chemical produced by a microorganism that kills or inhibits the

growth of another microorganism
• Antimicrobial agent: Chemical that kills or inhibits the growth of

microorganisms
Fleming and Penicillin
ANTIBIOTICS – Classification
• Accdg to antimicrobial activity 1. Bactericidal 2. Bacteriostatic
• Accdg to bacterial spectrum of activity 1. Narrow spectrum 2. Broad spectrum
• Accdg to mechanism of action
1. Inhibit bacterial cell wall synthesis
2. Alter the function and permeability of the cell membrane
3. Inhibit protein synthesis (translation and transcription)
4. Inhibit nucleic acid synthesis

1. Inhibit bacterial cell wall synthesis and
permeability of cell membrane
ß-Lactams
Cephalosporin Penicillin
• Cefalexin Narrow Spectrum
• Cefuroxime •Benzylpenicillin (Penicillin G)
• Cefotaxime •Phenoxymethylpenicillin (Pen V)
• Ceftriaxone
Broad Spectrum
Carbapenem •Amoxicillin/Co-amoxiclav
•Meropenem •Ampicillin
•Piperacillin with Tazobactam
•Imipenem
(Tazocin)
•Doripenem
•Ertapenem
3.Inhibit protein synthesis (translation and
transcription)
Aminoglycosides • Erythromycin
• Naturally occurring: • Azithromycin
• Streptomycin
• Neomycin
• Kanamycin
• Gentamicin
• Semisynthetic derivatives:
• Amikacin (from Kanamycin)
Macrolides
4. Inhibit nucleic acid synthesis
Flouroquinolones
• Ciprofloxacin
• Norfloxacin
• Moxifloxacin
• Levofloxacin
Uses of Antibiotic Sensitivity Testing
• Antibiotic sensitivity test: A laboratory test which determines how

effective antibiotic therapy is against a bacterial infections.
• Antibiotic sensitivity testing will control the use of antibiotics in

clinical practice
• Testing will assist the clinicians in the choice of drugs for the
treatment of infections.
Components of Antibiotic Sensitivity Testing
1.The identification of relevant pathogens in exudates and body fluids

collected from patients
2. Sensitivity tests done to determine the degree of sensitivity or resistance of

pathogens isolated from patient to an appropriate range of antimicrobial drugs
3. Assay of the concentration of an administered drug in the blood or body

fluid of patient required to control the schedule of dosage.
Components of Antibiotic Sensitivity Testing
• Drug-resistant bacteria have given rise to several serious outbreaks of infection,
with many deaths.
• This has led to a need for national and international surveillance programmes to
monitor antibiotic resistance in bacteria by susceptibility testing using reliable
methods that generate comparable data.
• The availability of microbiological and epidemiological information would help
clinicians in selecting the most appropriate antimicrobial agent for the treatment
of a microbial infection
Components of Antibiotic Sensitivity
Testing
• The susceptibility test must be performed by an accurate and
reproducible method, the results of which should be directly
applicable to the clinical situation.
• The ultimate criterion of the reliability of any susceptibility testing

method is its correlation with the response of the patient to
antimicrobial therapy.
General principles of antimicrobial susceptibility
testing
• Antimicrobial susceptibility tests measure the ability of an
antibiotic or other antimicrobial agent to inhibit bacterial
growth in vitro.
• This ability may be estimated by either the

• 1. dilution method or
• 2. the disc diffusion method
Dilution method
• This is for quantitative estimates of antibiotic activity.
• Dilutions of the antibiotic may be incorporated into broth or agar

medium, then inoculated with the test organism. Incubated overnight.
• The lowest concentration that prevents growth after overnight

incubation is known as the minimum inhibitory concentration (MIC)
of the agent.
Application of the MIC
• The MIC value obtained is then compared with known

concentrations of the drug obtainable in the serum and
in other body fluids to assess the likely clinical
response.
The disc diffusion test
• Paper discs are impregnated with an antibiotic.
• An agar medium is seeded uniformly with test

organism.
• The impregnated disc with the antibiotic, is placed on

the seeded agar medium with the test organism.
The disc diffusion test
• The antibiotic in the disc diffuses into the medium.
• A concentration gradient of the antibiotic forms by diffusion from the

disc. The growth of the test organism is inhibited at a distance from
the disc. This zone of inhibition is related, to the susceptibility of the
organism (among other factors).
Kirby-Bauer method
Interpretation of susceptibility test results
• Two category system
A--- susceptible
B--- Resistant
• Three category system

1. Susceptible
2. Intermediate susceptible
3. Resistant.
Two category system
• Susceptible
• An organism is called “susceptible” to a drug when the infection
it has caused is likely to respond to treatment with this drug, at
the recommended dosage.
• Resistant.
• The organism is expected not to respond to a given drug,
irrespective of the dosage and (also irrespective) of the location
of the infection.
Three category system
• Here in addition to susceptible and resistant

there is also “intermediate susceptible”.
Antimicrobial agent, Code Number %R %I %S
tested
Amoxicillin/Clavulanic acid, 816 75.6 6.9 16.4
AMC
Amikacin, AMK 1049 9 1.8 89.2
Ampicillin, AMP 1378 90.6 0.8 6.1
Azithromycin, AZM 2 50 0 50
Ceftazidime, CAZ 417 45.8 9.8 44.4

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Chemotherapy

• Chemotherapy refers to the use of drugs to treat a disease

• Antibiotic: Chemical produced by a microorganism that kills or inhibits the

• Antimicrobial agent: Chemical that kills or inhibits the growth of

• Accdg to antimicrobial activity 1. Bactericidal 2. Bacteriostatic

• Accdg to bacterial spectrum of activity 1. Narrow spectrum 2. Broad spectrum

• Accdg to mechanism of action

1. Inhibit bacterial cell wall synthesis

2. Alter the function and permeability of the cell membrane

3. Inhibit protein synthesis (translation and transcription)

4. Inhibit nucleic acid synthesis

• Antibiotic sensitivity test: A laboratory test which determines how

• Antibiotic sensitivity testing will control the use of antibiotics in

1.The identification of relevant pathogens in exudates and body fluids

2. Sensitivity tests done to determine the degree of sensitivity or resistance of

3. Assay of the concentration of an administered drug in the blood or body

• Drug-resistant bacteria have given rise to several serious outbreaks of infection,

with many deaths.

monitor antibiotic resistance in bacteria by susceptibility testing using reliable

methods that generate comparable data.

• The availability of microbiological and epidemiological information would help

• The ultimate criterion of the reliability of any susceptibility testing

• This ability may be estimated by either the

• This is for quantitative estimates of antibiotic activity.

• Dilutions of the antibiotic may be incorporated into broth or agar

• The lowest concentration that prevents growth after overnight

• The MIC value obtained is then compared with known

• Paper discs are impregnated with an antibiotic.

• An agar medium is seeded uniformly with test

• The impregnated disc with the antibiotic, is placed on

• The antibiotic in the disc diffuses into the medium.

• A concentration gradient of the antibiotic forms by diffusion from the

• Three category system

• Here in addition to susceptible and resistant

Amikacin, AMK 1049 9 1.8 89.2

Ampicillin, AMP 1378 90.6 0.8 6.1

Ceftazidime, CAZ 417 45.8 9.8 44.4

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