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Kirby-Bauer Disk Diffusion Susceptibility Test Send Print

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Created: Tuesday, 08 December 2009
Last update: Monday, 24 June 2013
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Kirby-Bauer disk diffusion susceptibility test
FIG. 1. Kirby-Bauer disk diffusion susceptibility test on coagulase-negative Staphylococcus aureus grown on Mueller-Hinton agar with tetracycline (30 µg), cephalothin (30 µg), erythromycin (15 µg), chloramphenicol (30 µg), vancomycin (30 µg), penicillin (10 µg), streptomycin (10 µg), and novobiocin (30 µg). (Tasha L. Sturm, Cabrillo College, Aptos, CA)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 2. Kirby-Bauer disk diffusion susceptibility test on Staphylocuccus aureus. The image depicts measuring the zone of inhibition for tetracycline. (Tasha L. Sturm, Cabrillo College, Aptos, CA)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 3. Kirby-Bauer disk diffusion susceptibility test on Pseudomonas aeruginosa. The results show sensitivity to amikacin and imipenem. Antibiotics used include ampicillin (A), cefotaxime (Ce), co-trimoxazole (Co), ciprofloxacin (Cf), amoxicillin-clavulanic acid (Ac), ceftazidime (Ca), amikacin (Ak), imipenem (I), and gentamicin (G). (Shashidhar Vishwanath, Kasturba Medical College, Karnataka, India)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 4. Kirby-Bauer disk diffusion test.  A Mueller-Hinton agar plate was seeded with a lawn of Pseudomonas aeruginosa using a sterile cotton swab.  Antibiotic disks containing 30 µg of tetracycline (upper left), 30 µg of vancomycin (upper right), 10 µg of ampicillin (lower left), and 30 µg of chloramphenicol (lower right) were dispensed on the agar surface, and the plate was incubated at 30°C overnight.  The diameter of each zone was measured in millimeters with a ruler and evaluated for susceptibility or resistance using the comparative standard method.  (Anh-Hue Tu, Georgia Southwestern State University, Americus)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 5. Kirby-Bauer disk diffusion test.  A Mueller-Hinton agar plate was seeded with a lawn of Pseudomonas aeruginosa (top half) and Serratia marcescens (bottom half) using sterile cotton swabs.  For plate A, antibiotic disks containing 30 µg of chloramphenicol (top and bottom left), 15 µg of erythromycin (top and bottom middle), and 30 µg of ampicillin (top and bottom right) were dispensed on the agar surface.  For plate B, antibiotic disks containing 25 µg of sulfisoxazole (top and bottom left) and 30 µg of ceftriaxone (top and bottom right) were dispensed on the agar surface.  Both plates were incubated at 30°C overnight and the diameter of each zone was measured in millimeters and evaluated for susceptibility or resistance using the comparative standard method. (Anh-Hue Tu, Georgia Southwestern State University, Americus)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 6. Kirby-Bauer disk diffusion test.  Mueller-Hinton agar plates were seeded with Pseudomonas aeruginosa, Serratia marcescens, and Staphylococcus aureus.  Four antibiotic disks were dispensed on each plate.  The disks contained 10 µg of ampicillin (top left), 30 µg of tetracycline (top right), 30 µg of chloramphenicol (bottom left), and 30 µg of vancomycin (bottom right).  All three plates were incubated at 30°C overnight.  The diameter of each zone was measured in millimeters and evaluated for resistance or susceptibility using the comparative standard method. Antibiotic susceptibility was compared between the three strains of bacteria. (Anh-Hue Tu, Georgia Southwestern State University, Americus)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 7. Kirby-Bauer disk diffusion test.  Mueller-Hinton agar plates were seeded with Pseudomonas aeruginosa and Staphylococcus aureus.  Four antibiotic disks were dispensed on each plate.  The disks contained 30 µg of tetracycline (top left), 30 µg of vancomycin (top right), 10 µg of ampicillin (bottom left), and 30 µg of chloramphenicol (bottom right).  Both plates were incubated at 30°C overnight.  The diameter of each zone was measured in millimeters and evaluated for susceptibility or resistance using the comparative standard method.  Antibiotic susceptibility was compared between the two strains of bacteria. (Anh-Hue Tu, Georgia Southwestern State University, Americus)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 8. Kirby-Bauer disk diffusion test.  Mueller-Hinton agar plates were seeded with Pseudomonas aeruginosa, Serratia marcescens, and Staphylococcus aureus.  Four antibiotic disks were dispensed on each plate.  The disks contained 30 µg of chloramphenicol (top left), 10 µg of ampicillin (top right), 30 µg of vancomycin (bottom left), and 30 µg of tetracycline (bottom right).  Both plates were incubated at 30°C overnight.  The diameter of each zone was measured in millimeters and evaluated for resistance or susceptibility using the comparative standard method. Antibiotic susceptibility was compared between the three strains of bacteria. (Anh-Hue Tu, Georgia Southwestern State University, Americus, GA)
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Kirby-Bauer disk diffusion susceptibility test

FIG. 9. Kirby-Bauer disk diffusion test.  A Mueller-Hinton agar plate was seeded with a lawn of Staphylococcus aureus using a sterile cotton swab.  Antibiotic disks containing 30 µg of ceftriaxone (upper left), 25 µg of sulfisoxazole (upper right), and 30 µg of polymixin B (lower right) were dispensed on the agar surface.  The plate was incubated at 30°C overnight.  The diameter of each zone was measured in millimeters with a ruler and evaluated for resistance or susceptibility using the comparative standard method. (Anh-Hue Tu, Georgia Southwestern State University, Americus, GA)

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Kirby-Bauer disk diffusion susceptibility test
FIG. 10. Kirby-Bauer disk diffusion test on Escherichia coli grown on Mueller-Hinton agar using antibiotic disks containing 10 µg of ampicillin (AM 10), 30 µg of tetracycline (Te 30), and 10 IU of penicillin (P 10) after a 24-hour incubation. (Jackie Peltier Horn, Houston Baptist University, Houston, TX)
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Kirby-Bauer disk diffusion susceptibility test

FIG. 11. Kirby-Bauer disk diffusion test. Oxacillin, doxycycline, and cefoxitin antibiotic disks were placed on Mueller-Hinton agar after plating with Staphylococcus aureus. The plate is shown just prior to incubation. (Clarissa L. Kaup, Bellevue University, Bellevue, NE; J.L. Henriksen, Bellevue University, Bellevue, NE)

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Kirby-Bauer disk diffusion susceptibility test

FIG. 12.  Kirby-Bauer disk diffusion test on Staphylococcus aureus grown on Mueller-Hinton agar. Zones of sensitivity are shown for oxacillin (15 mm), cefoxitin (30 mm), and doxycycline (27 mm) after 48 hours of incubation at 37°C. (Clarissa L. Kaup, Bellevue University, Bellevue, NE; J.L. Henriksen, Bellevue University, Bellevue, NE)

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Kirby-Bauer disk diffusion susceptibility test
FIG. 13. Kirby-Bauer disk diffusion test on Staphylococcus aureus grown on Mueller-Hinton agar. Staphylococcus aureus shows sensitivity to oxacillin (15-mm zone) after 48 hours of incubation at 37°C. (Clarissa L. Kaup, Bellevue University, Bellevue, NE; J.L. Henriksen, Bellevue University, Bellevue, NE)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 14. Kirby-Bauer disk diffusion test on Staphylococcus aureus grown on Mueller-Hinton agar.  Staphylococcus aureus shows decreased sensitivity to cefoxitin (11-mm zone) and doxycycline (26-mm zone) and resistance to oxacillin after 48 hours of incubation at 37°C. (Clarissa L. Kaup, Bellevue University, Bellevue, NE; J.L. Henriksen, Bellevue University, Bellevue, NE)
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Kirby-Bauer disk diffusion susceptibility test
FIG. 15. Kirby-Bauer disk diffusion test on Staphylococcus aureus grown on Mueller-Hinton agar. Staphylococcus aureus shows resistance to oxacillin after 48 hours of incubation at 37°C. (Clarissa L. Kaup, Bellevue University, Bellevue, NE, J.L. Henriksen, Bellevue University, Bellevue, NE)
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McFarland standards
FIG. 16. McFarland standards (left to right) 0.5, 1.0, 2.0, 3.0, positioned in front of a Wickerham card.  McFarland standards are used to prepare bacterial suspensions to a specified turbidity.  In the Kirby-Bauer disk diffusion susceptibility test protocol, the bacterial suspension of the organism to be tested should be equivalent to the 0.5 McFarland standard. (Jan Hudzicki, University of Kansas Medical Center, Kansas City, KS)
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Inoculation of the test plate
FIG. 17. Kirby-Bauer disk diffusion susceptibility test protocol, inoculation of the test plate. Step 2. Rotate the swab against the side of the tube while applying pressure to remove excess liquid from the swab prior to inoculating the plate. (Jan Hudzicki, University of Kansas Medical Center, Kansas City, KS)  
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Inoculation of the Mueller-Hinton agar plate
FIG. 18. Kirby-Bauer disk diffusion susceptibility test protocol, inoculation of the Mueller-Hinton agar plate. Step 3. Inoculate the plate with the test organism by streaking the swab in a back-and-forth motion very close together as you move across and down the plate.  Rotate the plate 60° and repeat this action.  Rotate the plate once more and repeat the streaking action.  This method ensures an even distribution of inoculum that will result in a confluent lawn of growth. (Jan Hudzicki, University of Kansas Medical Center, Kansas City, KS)
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Diagram illustrating the pattern the swab should follow as it is drawn across the plate
FIG. 19. Inoculation of the Mueller-Hinton agar plate, diagram illustrating the pattern the swab should follow as it is drawn across the plate. (Jan Hudzicki, University of Kansas Medical Center, Kansas City, KS)
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Inoculation of the Mueller-Hinton agar plate
FIG. 20. Kirby-Bauer disk diffusion susceptibility test protocol, inoculation of the Mueller-Hinton agar plate.  Step 4.  After streaking the Mueller-Hinton agar plate as described in Step 3, rim the plate with the swab by running the swab around the edge of the entire plate to pick up any excessive inoculum that may have been splashed near the edge.  The arrow indicates the path of the swab. (Jan Hudzicki, University of Kansas Medical Center, Kansas City, KS)

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