The susceptibility of CRKP isolates to 18 antibiotics, including amikacin (AK), penicillin (P), ampicillin (AMP), ampicillin/sulbactam (SAM), piperacillin (PIR), cefazolin (CFZ), cefaclor (CEC), cefuroxime (CFM), cefotaxime (CTX), ceftazidime (CAZ), ciprofloxacin (CIP), trimethoprim-sulfamethoxazole (SXT), fosfomycin (FOS), cefoperazone/sulbactam (SCF), cefepime (FEP), cefoxitin (FOX), aztreonam (ATM) and minocyline (MH), was determined by Kirby-Bauer disk diffusion test (Oxoid, United Kingdom). Five antibiotics, including imipenem (IMP), meropenem (MEM), ertapenem (ETP), tigecycline (TGC) and polymyxin B (PB), were tested by microbroth dilution test following the criteria of the Clinical and Laboratory Standards Institute (CLSI) (CLSI, 2016 ). The minimal inhibitory concentrations (MICs) breakpoint for tigecycline was defined according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) (EUCAST, 2017 ), while the others were interpreted according to CLSI protocols (CLSI, 2016 ). K. pneumoniae ATCC700603 and Escherichia coli ATCC25922 were used as quality control strains for the antibiotic susceptibility tests.
Kirby bauer disk diffusion test
Manufactured by Thermo Fisher Scientific
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The Kirby-Bauer disk diffusion test is a laboratory procedure used to determine the antibiotic susceptibility of bacterial isolates. The test involves placing antibiotic-impregnated disks on a culture plate inoculated with the bacterial sample, and then measuring the size of the zones of inhibition around the disks after incubation.
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3 protocols using kirby bauer disk diffusion test
1
Antibiotic Susceptibility of CRKP Isolates
2
Antibacterial Efficacy of Gelatin Hydrogels
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The susceptibility of S. aureus to s-gelatin, s-Cu-gelatin, and s-Zn-gelatin hydrogels was determined by methods similar to the Kirby-Bauer disk diffusion test (Oxoid, United Kingdom). Briefly, 100 μl of s-gelatin solution is mixed with or without 100 μl of 0.05 M copper or zinc solution to form hydrogel. S. aureus was planted on the Mueller-Hinton agar plate (MH plate) using a cotton swab, and different kinds of hydrogels were planted on the center of the MH plate. Moreover, the S. aureus planted in the MH plate without any intervene was labeled as blank control. All of the groups were incubated at 37°C with 5% CO2 for 48 hours. After the culture, the results were calculated and photographed, the approximate diameter of the antibacterial cycle was measured, and the statistical analysis was conducted according to the measured results.
3
Antibiotic Susceptibility of H. influenzae
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Antimicrobial susceptibility of H. influenzae isolates to 10 antibiotics, including ampicillin (AMP), ampicillin–sulbactam (AMS), ciprofloxacin (CIP), levofloxacin (LEV), ceftriaxone (CTR), cefuroxime (ROXH), cefotaxime (TAX), meropenem (MEM), trimethoprim–sulfamethoxazole (SXT), azithromycin (AZT), was determined by Kirby–Bauer disk diffusion test (Oxoid, United Kingdom) on Haemophilus test medium (HTM) plates and incubated for 24 h at 37 °C in air with 5% CO2. All the isolates were tested for the production of β-lactamase by nitrocephin paper disks.
Isolates that showed susceptibility to ciprofloxacin and had an inhibitory zone diameter of 21–28 mm were screened with nalidixic acid (NAL) disks (30 μg). Minimum inhibitory concentrations (MICs) of ciprofloxacin and levofloxacin were detected by E-test method (AB bioMerieux) in isolates resistant to ciprofloxacin or NAL disks. Susceptibility results were interpreted according to CLSI standards 26th Edition except NAL screening. NAL screening was interpreted according to EUCAST criteria [16 (link)]. H. influenzae was considered decreased susceptibility to ciprofloxacin when MICs of ciprofloxacin were ≥ 0.125 μg/ml. H. influenzae ATCC 29247 was used as a quality control strain for susceptibility testing.
Isolates that showed susceptibility to ciprofloxacin and had an inhibitory zone diameter of 21–28 mm were screened with nalidixic acid (NAL) disks (30 μg). Minimum inhibitory concentrations (MICs) of ciprofloxacin and levofloxacin were detected by E-test method (AB bioMerieux) in isolates resistant to ciprofloxacin or NAL disks. Susceptibility results were interpreted according to CLSI standards 26th Edition except NAL screening. NAL screening was interpreted according to EUCAST criteria [16 (link)]. H. influenzae was considered decreased susceptibility to ciprofloxacin when MICs of ciprofloxacin were ≥ 0.125 μg/ml. H. influenzae ATCC 29247 was used as a quality control strain for susceptibility testing.
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