For recognizing all the isolates, Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry (MALDI-TOF MS, Vitek MS, bioMérieux, France) was utilized. In accordance with the 2019 Clinical and Laboratory Standards Institute’s recommendations, 28 antibiotics were tested for their susceptibility via VITEK 2 (Card number: AST-GN13) system or Kirby-Bauer Disk Diffusion (Oxoid, UK) (CLSI) method. K. pneumoniae (ATCC700603) and Escherichia coli (ATCC25922) isolates were kept as standards to ensure quality control. Agar dilution assessment with the help of ceftazidime and cefotaxime combined with clavulanate was performed to confirm ESBL. Resistance to carbapenem (meropenem, imipenem, and ertapenem) was confirmed via disk diffusion protocol.
Kirby bauer disk diffusion
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The Kirby-Bauer Disk Diffusion test is a procedure used to determine the susceptibility of bacterial isolates to antimicrobial agents. The test involves placing antibiotic-impregnated disks on an agar plate inoculated with the bacterial culture. The zone of inhibition around each disk is then measured, providing information about the effectiveness of the antimicrobial agent against the specific bacterial strain.
Automatically generated - may contain errors
Lab products found in correlation
Vitek ms, by bioMérieux (1 mentions)
Kanamycin, by Thermo Fisher Scientific (1 mentions)
Nalidixic acid, by Thermo Fisher Scientific (1 mentions)
Ciprofloxacin, by Thermo Fisher Scientific (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Ampicillin, by Thermo Fisher Scientific (1 mentions)
Chloramphenicol, by Thermo Fisher Scientific (1 mentions)
Erythromycin, by Thermo Fisher Scientific (1 mentions)
Mueller hinton agar (mha), by Thermo Fisher Scientific (1 mentions)
Maldi tof ms, by Bruker (1 mentions)
Vitek 2, by bioMérieux (1 mentions)
4 protocols using kirby bauer disk diffusion
1
Comprehensive Antibiotic Susceptibility Profiling
2
Antimicrobial Susceptibility of E. coli
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Antimicrobial susceptibility testing of E. coli isolates was performed by the standard Kirby-Bauer disk diffusion method against selected antibiotics (Oxoid Ltd, Hampshire, United Kingdom) which includes; kanamycin (K 30 µg), nalidixic acid (NA 30µg), tetracycline (TE 30 µg), ampicillin (AMP 10µg), chloramphenicol (C 30 µg), erythromycin (E 15µg), gentamicin (CN 10 µg) and ciprofloxacin (CIP 5µg) on Mueller-Hinton agar (Oxoid Ltd, Hampshire, United Kingdom). The 2018 Clinical and Laboratory Standards Institute (CLSI) guidelines (13) were used for the interpretation and categorization of the test strains as sensitive (S), intermediate (I) or resistant (R). Control E. coli strain ATCC 25922 was also set along with positive isolates during susceptibility testing for each antibiotic used.
3
Characterization of Multidrug-Resistant Bacterial Strains
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Bacterial strains used in this study were methicillin-resistant Staphylococcus aureus ATCC 43300, Staphylococcus haemolyticus ATCC 29970 and Pseudomonas aeruginosa PAO1 as reference strains, and three multi-drug resistant clinical isolates: methicillin-resistant S. aureus (MRSA) 1118-116, methicillin and vancomycin-resistant S. haemolythicus (VRSH) 1219-118, and extended-spectrum beta-lactamase (ESBL) producing P. aeruginosa 0418-925. The strains were obtained from a collection previously established at the Department of Molecular Medicine and Medical Biotechnologies (University of Naples Federico II). No ethical approval was required for the study because there was no access to patients’ data. All strains were stored as 15% (v/v) glycerol stocks at −80 °C. Before each experiment, cells were sub-cultured from the stocks onto TSA plates at 37 °C for 24 h. Identification was performed by biochemical characterization using the Vitek2 (Biomerieux, Mercy-l’Etoile, France) and Phoenix (Becton Dickinson, Sparks, MD, USA) systems and confirmed by MS MALDI-TOF (Bruker Daltonics, Bremen, Germany). Susceptibility to antibiotics was assessed using automatic (Vitek2; Phoenix) and Kirby Bauer disk diffusion (Thermo Fisher Scientific, Basingstoke, UK) antibiotic sensitivity testing.
4
Antibiotic Susceptibility of M. catarrhalis
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The isolates were tested for susceptibility to erythromycin and azithromycin using the Kirby-Bauer disk diffusion (Thermo Fisher, Oxoid) method in accordance with the CLSI M45-A3 guideline (Clinical and Laboratory Standards Institute [CLSI], 2015 ). In addition, the E-test method was used to confirm the antimicrobial susceptibility results of macrolide-resistant M. catarrhalis. Staphylococcus aureus ATCC 25923 and ATCC 29213 were used for quality control.
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