Antibiotics Susceptibility

Test
Antibiotics Susceptibility Test
Two tests can be used: dilution series test and
agar diffusion test
Dilution series test antibiotic agents are
prepared in the nutrient medium, inoculated
with organism, incubated, then the lowest
growth-inhibiting concentration level are
determined
Very time-consuming
Kayser, Medical Microbiology 2005 Thieme
Antibiotics Susceptibility Test
Agar diffusion test inoculation of nutrient
agar plate with test strain, then disks of filter
paper containing antibiotic are placed upon
agar plate, incubated inhibition zone
around the disks (whether or not they
develop and their size) provide information
on the resistance of the microorganisms
tested
Kayser, Medical Microbiology 2005 Thieme
Minimal Inhibitory Concentration
MIC: the lowest concentration of drug that
inhibits the growth of the organism
The MIC is determined by inoculating the
organism isolated from the patient into a
series of tubes or cups containing 2-fold
dilutions of the drug dilution methods
After incubation at 35C for 18 hours, the
lowest concentration of drug that prevents
visible growth of the organism is the MIC
Disk diffusion method disks impregnated with
various antibiotics are placed on the surface of
an agar plate that has been inoculated with the
organism isolated from the patient
After incubation at 35C for 18 hours, during
which time the antibiotic diffuses outward from
the disk, the diameter of the zone of inhibition is
determined. The size of the zone of inhibition is
compared with standards to determine the
sensitivity of the organism to the drug.
Minimum Bactericidal Concentration
The minimum bactericidal concentration
(MBC): the smallest concentration of a
substance required to kill 99.9% of the
cells in an inoculum.
Determined using quantitative subcultures
from the macroscopically unclouded tubes
or (microplate) wells of an MIC dilution
series.
Kayser, Medical Microbiology 2005 Thieme
Determination of minimal inhibitory concentration (MIC) and minimal bactericidal
concentration (MBC). In the top part of the figure, the patient's organism is added to tubes
containing decreasing amounts of the antibiotic. After incubation at 37C overnight, growth
of the bacteria is observed visually.The lowest concentration of drug that inhibits growth,
ie, 3.1 pg/mL, is the MIC. However, at this point, it is not known whether the bacteria have
been killed or whether the drug has only inhibited their growth.To determine whether that
concentration of drug is bactericidal, ie, to determine its MBC, an aliquot (0.1 mL) from the
tubes is plated on an agar plate that does not contain any drug.The concentration of drug
that inhibits at least 99.9% of the bacterial colonies, ie, 6.2 pglmL, is the MBC.
Levinson W. Medical Microbiology and Immunology 8th Edition. Appleton & Lange.
1998.
Antibiotic sensitivity testing.A zone of inhibition surrounds several
antibiotic-containing disks. A zone of certain diameter or greater indicates
that the organism is sensitive.Some resistant organisms will grow all the
way up to the disk eg, disk N.
Levinson W. Medical Microbiology and Immunology 8th Edition. Appleton & Lange.
1998.
Antibiotics Susceptibility Test
This method, also known as the disk test, is used to test the
resistance of a bacterial culture to various anti-infective agents.
The method provides a basis for classification of a bacterial
strain as susceptible, resistant, or intermediate according to
the dimension of the inhibition zone.
Kayser, Medical Microbiology 2005 Thieme
Antibiotics Susceptibility Test
Each point represents a bacterial strain. The size of the inhibition zone is
determined in the agar diffusion test (disk test) and the minimum inhibitory
concentration (MIC) in the dilution test. The MIC corresponds logarithmically (log2)
with the diameter of the inhibition zone.
Kayser, Medical Microbiology 2005 Thieme
Antibiotics Susceptibility Test
Interpretation of the results: the MICs or inhibition zones
are brought into relation with the substance
concentrations present at a site of infection at standard
dosage levels
This calculation is based on various pharmacokinetic
(serum concentration, half-life) and pharmacodynamic
parameters (bactericidal activity or not, postantibiotic
effect, etc.)
Also based on clinical experience gained from therapy of
infections with pathogens of given suceptibility.
Such data are used to establish general guideline values
defining the boundary between susceptible and resistant
bacteria
Kayser, Medical Microbiology 2005 Thieme