Disk diffusion tests (Oxoid, Hampshire, United Kingdom) were performed according to EUCAST recommendations version 7.0 (2019). MH agar plates were supplemented with 50 µg/mL kanamycin, 250 ng/mL of the inducer anhydrotetracycline, and/or 50 µg/mL cloxacillin (class C β-lactamase inhibitor). Cultures were incubated at 37 °C for 24 h before measuring inhibition zones. Each disk of cefotaxime, ceftazidime, cefoxitin, cefepime, and aztreonam contained 30 µg of the antibiotic. Ertapenem disks contained 10 µg, and the amoxicillin–clavulanate combination disk contained 30 µg of the antibiotic and 10 µg of the inhibitor. Potential inhibition of the β-lactamase by clavulanic acid was controlled in a double disk synergy test by placing the cefotaxime and the amoxicillin–clavulanate combination disk at a distance of 20 mm center to center.
Disk diffusion tests
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, China
Disk diffusion tests are a standardized method for determining the susceptibility of bacteria to antimicrobial agents. The tests involve placing antibiotic-impregnated disks on a bacterial culture lawn and measuring the zone of inhibition around each disk, which indicates the sensitivity of the bacteria to the tested antimicrobials.
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2 protocols using disk diffusion tests
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Antibiotic Disk Diffusion Testing
2
Antimicrobial Susceptibility Testing: Disk Diffusion
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Antimicrobial susceptibility testing was performed by using the disk diffusion tests (Oxoid) on Mueller Hinton agar (KeMaJia, China) according to Clinical and Laboratory Standards Institute guidelines (Wayne, 2015) , with the following antibiotics being included: ampicillin (10 mg), piperacillin (100 mg), amikacin (30 mg), piperacillin-tazobactam (100 mg/10 mg), trimethoprimsulfamethoxazole (1.25 mg/23.75 mg), levofloxacin (5 mg), ciprofloxacin (5 mg), gentamicin (10 mg), ampicillin-sulbactam (10 mg/ 10 mg), ceftazidime (30 mg), cefepime (30 mg), chloramphenicol (30 mg), imipenem (10 mg). E.coli ATCC25922 was used as the quality control in parallel.
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