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Antimicrobial Susceptibility Testing

This document discusses antimicrobial susceptibility testing (AST), which determines the levels of resistance or susceptibility of bacterial isolates to antibiotics. It describes the disk diffusion (Kirby-Bauer) method, where disks impregnated with antibiotics are placed on inoculated agar plates to measure inhibition zone diameters. The minimum inhibitory concentration (MIC) tube dilution test determines the lowest antibiotic concentration that prevents bacterial growth. Results are interpreted as susceptible, intermediate, or resistant to guide appropriate antibiotic treatment. Standardization and automation help ensure accuracy and efficiency of AST.

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Fatimah Tambilawan
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303 views14 pages

Antimicrobial Susceptibility Testing

This document discusses antimicrobial susceptibility testing (AST), which determines the levels of resistance or susceptibility of bacterial isolates to antibiotics. It describes the disk diffusion (Kirby-Bauer) method, where disks impregnated with antibiotics are placed on inoculated agar plates to measure inhibition zone diameters. The minimum inhibitory concentration (MIC) tube dilution test determines the lowest antibiotic concentration that prevents bacterial growth. Results are interpreted as susceptible, intermediate, or resistant to guide appropriate antibiotic treatment. Standardization and automation help ensure accuracy and efficiency of AST.

Uploaded by

Fatimah Tambilawan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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LICEO DE CAGAYAN

UNIVERSITY COLLEGE OF MEDICAL


LABORATORY SCIENCE CLINICAL BACTERIOLOGY
LABORATORY

ANTIMICROBIAL
SUSCEPTIBILITY
TESTING
PRESENTED BY:
JOHN PATRICK B. TUDAS, RMT
❖Identification of a bacterial isolate from a patient provides guidance in the
choice of an appropriate antibiotic for treatment

❖Many bacterial species are not uniformly susceptible to a particular


anti-bacterial compound

❖This is particularly evident among the Enterobacteriaceae, Staphylococcus


spp., and Pseudomonas spp.

❖The wide variation in susceptibility and high frequencies of drug resistance


among strains in many bacterial species necessitates the determination of
levels of resistance or susceptibility as a basis for the selection of the
proper antibiotic for chemotherapy
SELECTION OF ANTIBIOTICS
❖Generally, labs choose 10-15 antibiotics to test susceptibility
for GP organisms and another 10-15 for GN organisms.

❖Each laboratory should have a battery of antibiotics ordinarily


used for testing

❖Drug formulary decided by medical staff, pharmacists, and


medical technologists
IDEAL ANTIMICROBIAL AGENT

1. highly toxic to the microbe;


2. non-toxic to the host;
3. not interfere with the ability of the host to fight
other diseases; and
4. not lead to the development of drug resistance.
STANDARDIZATION OF AST

❖ Inoculum Preparation
• Use 4-5 colonies NOT just 1 colony
• Saline or Broth

❖ Inoculum Standardization
• using 0.5 McFarland standard (1.5 X
10^8 CFU/ml)
DISK-DIFFUSION METHOD (KIRBY-BAUER METHOD):

❖ The disk-diffusion method (Kirby-Bauer) is


more suitable for routine testing in a clinical
laboratory where a large number of isolates
are tested for susceptibility to numerous
antibiotics.

❖ An agar plate is uniformly inoculated with the


test organism
❖ A paper disk impregnated with a fixed
concentration of an antibiotic is placed on the agar
surface.
DISK-DIFFUSION METHOD (KIRBY-BAUER METHOD):

❖ Growth of the organism and diffusion of the


antibiotic commence simultaneously resulting in
a circular zone of inhibition in which the amount
of antibiotic exceeds inhibitory concentrations.

❖ The diameter of the inhibition zone is a function of


the amount of drug in the disk and susceptibility
of the microorganism.
MUELLER-HINTON AGAR (MHA) PLATE
MINIMUM INHIBITORY CONCENTRATION (MIC)
❖ The tube dilution test is the standard method for
determining levels of resistance to an antibiotic.

❖ Serial dilutions of the antibiotic are made in a liquid


medium which is inoculated with a standardized
number of organisms and incubated for a prescribed
time.

❖ The lowest concentration of antibiotic preventing


appearance of turbidity is considered to be the minimal
inhibitory concentration (MIC).
E TEST / GRADIENT DIFFUSION TEST
MINIMUM INHIBITORY CONCENTRATION (MIC)
❖ Susceptible ”S”
• Interpretive category that indicates an organism is inhibited by
the recommended dose, at the infection site, of an
antimicrobial agent

❖ Intermediate “I”
• Interpretive category that represents an organism that may
require a higher dose of antibiotic for a longer period of time to be
inhibited

❖ Resistant “R”
• Interpretive category that indicates an organism is not inhibited
by the recommended dose, at the infection site, of an
antimicrobial agent.
AUTOMATION OF AST

❖ Detect growth in microvolumes of broth with


various dilutions of antimicrobials
❖ Detection via photometric, turbidimetric, or
fluorometric methods
▪ Types
1. BD Phoenix
2. Microscan Walkaway
3. TREK Sensititre
4. Vitek 1 and 2
LICEO DE CAGAYAN
UNIVERSITY COLLEGE OF MEDICAL
LABORATORY SCIENCE CLINICAL BACTERIOLOGY
LABORATORY

END OF
DISCUSSION

PRESENTED BY:
JOHN PATRICK B. TUDAS, RMT

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