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Kirby Bauer Disc Diffusion Method For Antibiotic
Susceptibility Testing
May 18, 2022 by Reeju Sharma
Edited By: Sagar Aryal
As the Broth dilution method are time‐consuming, many laboratories in the United
States adopted disc diffusion tests in the early 1950s. Kirby Bauer tests also known as
the Disc diffusion test is used for antibiotic susceptibility testing. Lack of
standardization creates a problem in the 1960s and later Kirby and Bauer reviewed
the description. In 1961, WHO standardized the procedure. Currently, CLSI updates
and modifies the original procedure which ensures uniformity worldwide. This test is
mainly performed to determine the sensitivity or resistivity of aerobes or facultatively
anaerobes against different classes of antibiotics.
Kirby Bauer Disc Diffusion Method For Antibiotic Susceptibility Testing. Created with BioRender.com
Table of Contents
Table of Contents
Aim/ Objectives
Principle of Kirby Bauer Disc Diffusion Method
Materials Required
Procedure of Kirby Bauer Disc Diffusion Method
Result and Interpretation of Kirby Bauer Disc Diffusion Method
Uses of Kirby Bauer Disc Diffusion Method
Advantages
Limitations
References
Aim/ Objectives
This technique aids physicians to assist in selecting treatment options.
The growth of organisms determines the ability of antibiotics to inhibit the
organisms.
Principle of Kirby Bauer Disc Diffusion Method
The organism to be tested must be incubated overnight in broth and must be
compared with the 0.5 McFarland Turbidity standard. Mueller‐ Hinton agar must be
used as it does not inhibit sulphonamides and ensures reproducibility and with
composition and PH of the medium. The agar when poured on Petri dishes should be
4mm.
Materials Required
Mueller‐ Hinton agar
Antibiotic discs
Cotton swabs
Petri dishes
0.5 McFarland Turbidity standard
Inoculum
Forceps
Metric ruler or caliper
Procedure of Kirby Bauer Disc Diffusion Method
1. Sterilize the area with disinfectant and open burner before performing the test.
2. A sterile cotton swab is dipped into the inoculum and remove excess medium by
pressing the swab onto the wall of the tube.
3. Swab the surface area of the plate completely by rotating the plate. This technique
is called lawn culture or carpet culture.
4. Allow the plates to dry for 5 minutes so that the medium absorbs the inoculum
properly.
5. Firstly sterilize the forceps with alcohol before picking up antibiotic discs.
6. Discs should be placed at a distance of 24mm.
7. Lightly touch each disc with forceps to ensure that it is in good contact to avoid
misplacement.
8. Incubate the plate upside down for 24 hours at 37ºC.
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Result and Interpretation of Kirby Bauer Disc Diffusion Method
After 24 hours of incubation, use a metric ruler to measure the zone of inhibition
and include the diameter of the disc in the measurement.
Compare the result with CLSI guidelines to report the result.
The results are reported as Susceptible ﴾S﴿, Intermediate ﴾I﴿, or Resistant ﴾R﴿.
Uses of Kirby Bauer Disc Diffusion Method
Antibiotic susceptibility testing is a very important step to monitor antimicrobial
resistance.
It guides the clinician to select the best antimicrobial agent.
To acquire information on microorganisms of public health importance.
Advantages
This test is used in determining the antibiotics of choice to treat an infection.
It can be useful in monitoring antimicrobials and for the selection of proper
antibacterial agents.
It doesn’t require special equipment to perform and can be interpreted by all
medical personnel.
It costs less to perform this test.
Limitations
Not all slow or fastidious organisms can be tested accurately.
It is not considered a gold standard test as it is only for screening the susceptibility
pattern of the organisms according to the CLSI guideline.
Read Also:
Microbial Culture Media‐ Definition, Types, Examples, Uses
Serial Dilution: Formula, Calculator, Procedure, Uses
22 Types of Spectroscopy with Definition, Principle, Steps, Uses
Bacteria‐ Definition, Structure, Shapes, Sizes, Classification
Antimicrobial Susceptibility Testing ﴾AST﴿‐ Types and Limitations
References
https://en.wikipedia.org/wiki/Antibiotic_sensitivity_testing – 5%
https://www.asmscience.org/content/education/protocol/protocol.3189 – 2%
https://asm.org/getattachment/2594ce26‐bd44‐47f6‐8287‐0657aa9185ad/Kirby‐
Bauer‐Disk‐Diffusion‐Susceptibility‐Test‐Protocol‐pdf.pdf – 1%
https://www.pharmadekho.com/swab‐testing‐of‐various‐surfaces‐for‐bioburden‐
determination/ – 1%
https://www.researchgate.net/publication/279912715_Current_Methods_for_the_Id
entification_of_Carbapenemases – 1%
About Author
Reeju Sharma
Reeju Sharma completed her Master’s degree ﴾M.Sc.﴿ in medical microbiology from
St. Xavier's College, Tribhuvan University, Kathmandu, Nepal, and did her
bachelor’s degree ﴾B.Sc.﴿ in general microbiology. She is currently working in a
pharmaceutical company as a Quality control officer.
3 thoughts on “Kirby Bauer Disc Diffusion Method For Antibiotic
Susceptibility Testing”
TIMOTHY KIBII
November 14, 2022 at 7:15 PM
very useful
Reply
Stephen otieno
May 13, 2022 at 3:10 PM
Good notes for a concerned student
Reply
Stephen otieno
May 13, 2022 at 3:09 PM
Nice information source asit gives appreciatable answers though.
Reply
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