Antibiotic

sensitivity testing

 Used to select effective antimicrobial

drugs for bacterial pathogens.
Must never be performed on
commensals/contaminants.
Only measures antibiotic sensitivity in
vitro, NOT IN VIVO.

 When selecting antimicrobial treatment, the

doctor should:
Consider the patients clinical condition
Type and site of infection.
History of drug hypersensitivity
Activity of different drugs i.e. rate of absorption,
diffusion in tissues, metabolism, excretion and
toxicity and effects on patients normal microbial
flora.

Antimicrobials may occur as:

Natural products
Semisynthetic products
Synthetic chemicals.

Antimicrobial agents can be:

Bacteriostatic
Bactericidal
Classified by their range of activities
Narrow spectrum antibiotics
Broad spectrum antibiotics
Family, organ system on which they exert
their action, and by their mode of action.

Mechanisms of antimicrobial action effect:

Microbial cell wall and membrane.
Microbial protein synthesis.
Nucleic acid synthesis.
Synthesis of metabolites necessary for
growth.

Antibiotic sensitivity testing in the clinical

laboratory provides the clinician vital
information for the selection of the most
appropriate antibiotic to be used for the
specific infection.
Microbial sensitivity to therapeutic doses of
specific antibiotics is indicated by clear zones
around the antibiotic discs.

The Kirby-Bauer procedure is an antimicrobial

susceptibility
Susceptibility (sensitive) implies the organism
should respond favourable to therapeutic
doses of the drug.
Intermediate implies some susceptibility to
therapeutic doses.
Resistance the usual therapeutic doses
would be ineffective.

Sensitivity testing techniques:

1. Diffusion method
Kirby-Bauer method
Stokes diffusion method
2. Dilution technique
Macrodilution broth susceptibility test
Agar dilution method

Turbidity [opacity] standard

This is a barium chloride standard
against which the turbidity of the test and
control inocula can be compared.
When matched with the standard, the
inocula should give a semi confluent
growth.

Indirect/Direct sensitivity testing

Is when a pure culture of the pathogen is used as the
inoculum
Direct/Primary sensitivity testing
Performed on specimens such as positive blood
culture, CSF showing only one type of organism on the
gram stain/other specimens showing large numbers of
one type of organism
Results of direct test to confirmed by the indirect test.

 A set of round I antibiotics are used first.

A set of round II of antibiotics are used if
and only when one antibiotic is sensitive
or when all antibiotics are resistant.

Factors which can affect reading of sensitivity tests

Sensitivity test media too thick

Sensitivity test media too thin

Avoid picking more than one colony if there is mixed

growth of organisms.

Check that all materials used are sterile.

Use the prescribed discs for the organism tested

Use discs of correct antimicrobial content

Always measure zone sizes that are less than control

If growth is not semi confluent/there are signs of

mixed growth on sensitivity plate-repeat test.