Antibiotic Sensitivity Testing
•Antibiotic sensitivity testing or antibiotic
susceptibility testing is the measurement of
the susceptibility of bacteria to antibiotics.
•Sensitivity testing results can allow a clinician
to change the choice of antibiotics from empiric
therapy, which is when an antibiotic is selected
based on clinical suspicion about the site of an
infection and common causative bacteria ,to
directed therapy, in which the choice of antibiotic
is based on knowledge of the organism and its
sensitivities.
•Sensitivity testing usually occurs in a medical
laboratory, and it can be done by four methods.
• Method One :-
The isolated organism from the patient is inoculated in
a series of tubes containing 2-fold dilutions of the drug
and incubation at 37°C for 18 hours.
The lowest drug concentration that prevents visible
growth is the Minimal Inhibitory Concentration (MIC).
This provides the physician with the precise
concentration of drug to guide the choice of both the
drug and the dose.
• Method two:-
- Disk Diffusion method, in which disks impregnated
with various antibiotics are placed on the surface of an
agar plate that has been inoculated with the organism
isolated form the patient.
- After incubation at 37°C 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.
• Method Three:-
•Genetic methods such as polymerase chain reaction
(PCR) that test to see if bacteria have genes that confer
resistance.
• Method Four:-
E - test (known as the Epsilometer test)is a way
of determining antimicrobial sensitivity by
placing a strip impregnated with antimicrobials
onto an agar plate.
A strain of bacterium or fungus will not grow
near a concentration of antibiotic or antifungal
if it is sensitive.
It can be used to determine a minimum
inhibitory concentration (MIC).
Minimal Inhibitory Concentration (MIC): The lowest
concentration of drug that inhibits the growth of the
organism.
Minimal Bactericidal Concentration (MBC): The
concentration of drug that actually kills the organism, not
just merely inhibiting growth.
• It must be determined for certain infections such as
endocarditis, by taking a small sample ( 0.01 or 0.1 mL)
from the tube used for the MIC assay and spreading it
over the surface of a drug-free blood agar plates.
• Any organisms that were inhibited but not killed now
have a chance to grow because the drug has been
diluted significantly.
• After incubation for 48 hours at 37°C, the lowest
concentration that has reduced the number of colonies
by 99.9% compared with the drug-free control is the MBC.
Determination of the MIC and the MBC.
- Bactericidal drugs usually have an MBC equal or very
similar to the MIC, whereas bacteriostatic drugs usually
have an MBC significantly higher than the MIC.
Antibiotic Sensitivity Testing.
E-test being used to determine the susceptibility of
Neisseria gonorrhoeae to benzylpenicillin.
Use of Antibiotic Combinations
In most cases, the single best antimicrobial
agent should be selected for use because this
minimizes side effects.
However, there are several instances in which
two or more drugs are commonly given:-
1- To treat serious infections before the identity
of the organism is known.
2- To achieve a synergistic inhibitory effect
against certain organisms.
3- To prevent the emergence of resistant
organisms.
Two drugs can interact in one of several ways:-
1- Indifference: This is when the two drugs are additive
to each other.
For Example, the combination of a penicillin with an
aminoglycoside.
2- Synergism, in which the effect of the two drugs
together is significantly greater than the sum of the
effects of the two drugs acting separately.
For Example, the combination of a sulfonamide and
trimethoprim.
3- Antagonism, in which the result is significantly lower
activity than the sum of the activities of the two drugs
alone.
For Exapmle, using penicillin G with tetracycline.
Drug interactions
Genetic Basis of Resistance
1- Chromosomal mutations: Change the target of the
drug or change the membrane. They occur at a low
frequency (perhaps 1 in 10 million organisms), affect
only one drug.
2- Resistance plasmids (R factors): Usually carry 2
sets of genes. One set encodes enzymes that degrade
or modify drugs and the other encodes proteins that
mediate cojugation, the main process by which
resistance genes are transferred from one bacterium
to another. Plamid-mediated resistance occurs at a
higher frequency affecting multiple drugs.
3- Transposons are small pieces of DNA that move
from one site on the bacterial chromosome to another
or from bacterial chromosome to plasmid DNA.
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