Antimicrobial susceptibility testing was performed using the Kirby–Bauer disc diffusion method against the following antibiotic discs according to the Clinical and Laboratory Standards Institute guidelines [28 ]. Microbial susceptibility test was conducted using the following antimicrobials: gentamicin (10 μg), tobramycin (10 μg), amikacin (30 μg), imipenem (10 μg), meropenem (10 μg), ciprofloxacin (5 μg), ceftazidime (30 μg), piperacillin/tazobactam (100/10 μg), ampicillin/sulbactam (10/10 μg) and levofloxacin (5 μg) (Mast Group Ltd., Bootle Merseyside, UK). Acinetobacter baumannii ATCC 19606 was used as the quality control strain in antimicrobial susceptibility testing.
Tobramycin
Manufactured by Mast Group
Sourced in United Kingdom
Tobramycin is a laboratory-grade antibiotic compound used in research and development applications. It is a water-soluble powder that can be dissolved in appropriate solvents for various experimental purposes.
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Lab products found in correlation
Amikacin, by Mast Group (9 mentions)
Gentamicin, by Mast Group (7 mentions)
Ciprofloxacin, by Mast Group (7 mentions)
Imipenem, by Mast Group (6 mentions)
Ceftazidime, by Mast Group (5 mentions)
Piperacillin tazobactam, by Mast Group (4 mentions)
Ceftriaxone, by Mast Group (4 mentions)
Meropenem, by Mast Group (3 mentions)
Levofloxacin, by Mast Group (3 mentions)
Tetracycline, by Mast Group (3 mentions)
Trimethoprim sulfamethoxazole, by Mast Group (3 mentions)
Piperacillin, by Mast Group (3 mentions)
Cefepime, by Mast Group (3 mentions)
Ampicillin sulbactam, by Mast Group (2 mentions)
Rifampicin, by Mast Group (2 mentions)
Kanamycin, by Mast Group (2 mentions)
Linezolid, by Mast Group (2 mentions)
Teicoplanin, by Mast Group (2 mentions)
Mueller hinton agar (mha), by Merck Group (2 mentions)
Quinupristin dalfopristin, by Mast Group (2 mentions)
9 protocols using tobramycin
1
Antimicrobial Susceptibility Testing of Acinetobacter baumannii
2
Methicillin Resistance Screening in S. aureus
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The disk diffusion method using cefoxitin (30 μg) disk in Mueller-Hinton agar (Merck, Germany) according to the CLSI was applied for the screening of methicillin resistance isolates. In addition, PCR assay was used for the detection of mecA gene.12
The Kirby–Bauer disk diffusion method was used to determine the susceptibility of the isolates against penicillin, ceftriaxone, amikacin, gentamicin, tobramycin, kanamycin, tetracycline, linezolid, teicoplanin, ciprofloxacin, rifampicin, quinupristin-dalfopristin, and trimethoprim-sulfamethoxazole (Mast Co., UK) based on the CLSI recommendation (CLSI 2019). The minimum inhibitory concentration (MIC) value for vancomycin, mupirocin, tigecycline, and fusidic acid was determined using the broth microdilution method. Results for fusidic acid and tigecycline were interpreted according to the European Committee for antimicrobial susceptibility testing (EUCAST) breakpoints (http://www.eucast.org
The Kirby–Bauer disk diffusion method was used to determine the susceptibility of the isolates against penicillin, ceftriaxone, amikacin, gentamicin, tobramycin, kanamycin, tetracycline, linezolid, teicoplanin, ciprofloxacin, rifampicin, quinupristin-dalfopristin, and trimethoprim-sulfamethoxazole (Mast Co., UK) based on the CLSI recommendation (CLSI 2019). The minimum inhibitory concentration (MIC) value for vancomycin, mupirocin, tigecycline, and fusidic acid was determined using the broth microdilution method. Results for fusidic acid and tigecycline were interpreted according to the European Committee for antimicrobial susceptibility testing (EUCAST) breakpoints (
3
Antibiotic Resistance Profiling of Bacterial Isolates
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The resistance pattern of isolates against 15 antibiotics including 5 fluoroquinolones was determined by disc diffusion method on Mueller-Hinton agar (Merck, Germany) as described by the Clinical Laboratory Standards Institute (CLSI 2017) guidelines36 . The antibiotic disks used were as follows: amikacin (30 μg), aztreonam (30 μg), cefepime (30 μg), ceftazidime (30 μg), ceftriaxone (30 μg), ciprofloxacin (5 μg), gatifloxacin (5 μg), norfloxacin (5 μg), gentamycin (10 μg), imipenem (10 μg), levofloxacin (5 μg), ofloxacin (5 μg), piperacillin (100 μg), piperacillin/tazobactam (100 μg/10 μg), and tobramycin (10 μg) (MAST Co., Berkshire, UK). Drug-resistant patterns were defined as follows: MDR isolates (resistant to at least three antibiotics belonging to different chemical classes), XDR strains (resistant to at least one agent in all but two or fewer antimicrobial groups), and PDR strains (resistant to all antimicrobial classes)37 (link). E. coli ATCC 25922 and P. aeruginosa ATCC 27853 were used as quality control strains.
4
Comprehensive Antibiotic Susceptibility Testing
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The Kirby–Bauer disc-diffusion method on Mueller–Hinton agar was used to test the susceptibility of the isolates against amikacin, gentamicin, tobramycin, kanamycin, tetracycline, erythromycin, clindamycin, linezolid, teicoplanin, ciprofloxacin, rifampicin, quinupristin-dalfopristin and trimethoprim-sulfamethoxazole (Mast Co., Bootle, UK) based on the CLSI guideline. Susceptibility to vancomycin, mupirocin, tigecycline and fusidic acid was assessed by the broth microdilution method to determine the MIC titre. The European Committee for Antimicrobial Susceptibility Testing (EUCAST) breakpoints was used to determine MIC titres of fusidic acid and tigecycline (EUCAST 2018). The results of other antibiotics were interpreted by using the CLSI 2018 breakpoints. Low-level and high-level mupirocin resistance (LLMUPR, HLMUPR), inducible macrolide-lincosamide-streptogramin group B (iMLSB) and constitutive (cMLSB) macrolide-lincosamide-streptogramin group B were identified based on the CLSI guideline. Susceptibility testing was quality controlled using S. aureus ATCC 25923, ATCC 43300 and ATCC 29213 strains. Powders of antibiotics were all obtained from Sigma Chemical Co. (St Louis, MO, USA).
5
Antimicrobial Susceptibility Testing for P. aeruginosa
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The antimicrobial susceptibility test was evaluated using the Kirby Bauer disk diffusion method and interpreted based on the Clinical Laboratory Standard Institute (CLSI). The antibiotic disks included Imipenem (10 μg), Ceftazidime (30 μg), Piperacillin (100 μg), Gentamicin (10 μg), Amikacin (30 μg), Ciprofloxacin (5 μg), and Tobramycin (10 μg) (MAST Co., England). In this study, P.aeruginosa ATCC 27853 was used as a control strain.
6
Antimicrobial Susceptibility of Lactobacillus
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Qualitative antibacterial susceptibility of the microorganisms was determined according to the standard disk diffusion (Kirby-Bauer) method (20 (link)). using the paper disk including (μg/disc): azithromycin (15 (link)); ceftriaxone (21 (link)); imipenem (10 (link)); amikacin (21 (link)); chloramphenicol (21 (link)); ceftazidim (21 (link)); tobramycin (10 (link)); gentamycin (10 (link)) andciprofloxacin (5 (link)) purchased from Mast Co (Liverpool, UK).
Microbial suspensions with 106 colony forming units (CFU/mL) of each Lactobacillus strain in NB were prepared on a De Mann Rogosa Sharpe agar plate. The plates were incubated at 37°C for 48 hours in anaerobic condition and examined for the inhibition zone diameter appearing around each antibiotic disc. A test was carried out thrice for each antibiotic agent . Inhibitory zone diameters were compared with the standards provided by the National Committee for Clinical Laboratory Standards (NCCLS) (19 ). Antibiotic sensitivity of P. aeruginosa was also determined by the same method using the Muller Hilton agar.
Microbial suspensions with 106 colony forming units (CFU/mL) of each Lactobacillus strain in NB were prepared on a De Mann Rogosa Sharpe agar plate. The plates were incubated at 37°C for 48 hours in anaerobic condition and examined for the inhibition zone diameter appearing around each antibiotic disc. A test was carried out thrice for each antibiotic agent . Inhibitory zone diameters were compared with the standards provided by the National Committee for Clinical Laboratory Standards (NCCLS) (19 ). Antibiotic sensitivity of P. aeruginosa was also determined by the same method using the Muller Hilton agar.
7
Microbial Contamination of ATM Keypads
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Out of 360 ATMs at four locations in Hamadan (number provided by the central bank of the islamic republic of Iran) 96 ATMs were randomly selected. Using a sterile swab soaked in saline, samples were taken from the surfaces of the ATM keyboards. The swab was put into nutrient broth media and transferred to the microbiology laboratory of the University of Medical Sciences, Hamadan and incubated for 30 min. In order to differentiate microorganisms, swabs were cultured on blood agar and MacConkey agar plates and incubated at 37°C for 18–24 hours. Identification of the isolated bacteria was performed using standard microbiological methods [16 ]. For all isolated strains, antibacterial susceptibility was tested using the standard Kirby-Bauer disk agar diffusion (DAD) method on Mueller Hinton agar (Merk Co., Germany) according to the clinical and laboratory standards institute guidelines (CLSI; 2015, M100-S25) [17 (link)] using gentamicin (10 µg), vancomycin (30 µg), trimethoprim/sulfamethoxazole (25 µg), amikacin (30 µg), tobramycin (10 µg), cephalotin (30 µg), norfloxacin (5 µg), and ceftizoxim (30 µg) disks (Mast Co.UK).
All statistical analyses were performed using the SPSS software package, version 21.
All statistical analyses were performed using the SPSS software package, version 21.
8
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility testing was carried out by the Kirby–Bauer disc diffusion method according to Clinical and Laboratory Standards Institute (CLSI) [12 ] and breakpoint interpretations for including amikacin (30 µg), gentamicin (10 µg), tobramycin (10 µg), cotrimoxazole (trimethoprim–sulfamethoxazole) (25 µg), tigecycline (15 µg), doxycycline (30 µg), tetracycline (30 µg), minocycline (30 µg) ceftriaxone (30 µg), ceftazidime (30 µg), cefepime (30 µg), cefotaxime (30 µg), piperacillin/tazobactam (100/10 µg), ampicillin-sulbactam (10/10 µg), imipenem (10 µg), meropenem (10 µg), ciprofloxacin (5 µg), levofloxacin (5 µg), (Mast Group Ltd., Bootle, UK). The minimum inhibitory concentrations (MICs) of colistin (Colistin sulfate salt powder, Sigma-Aldrich, St. Louis, MO, USA) were determined using the broth microdilution method according to CLSI guidelines (CLSI, 2020). Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27,853 strains were used as the standard strains.
9
Antimicrobial Susceptibility Testing for Bacterial Isolates
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Antimicrobial susceptibility testing for the bacterial isolates was carried out using the Kirby-Bauer method, as recommended by the Clinical and Laboratory Standards Institute (CLSI). 15 The following antibiotics were tested: imipenem (10 μg), meropenem (10 μg), doripenem (10 μg), ceftazidime (30 μg), cefepime (30 μg), piperacillin (100 μg), piperacillin/tazobactam (100/10 μg), gentamicin (10 μg), amikacin (30 μg), tobramycin (10 μg), ciprofloxacin (5 μg), aztreonam (30 μg), polymyxin B (300 units), and colistin (10 μg) (Mast Group Ltd, UK). The minimum inhibitory concentrations (MICs) of carbapenems (imipenem [IMI], meropenem [MRP], and doripenem [DOR]) were obtained by an E-test (Liofilchem, Italy), as described in the manufacturer's instructions. Carbapenem resistance was determined based on the MIC breakpoints. When an isolate was resistant to three carbapenems (imipenem, meropenem, and doripenem), that isolate was considered high-level carbapenem resistant. If an isolate was resistant to three or more classes of antimicrobial agents (i.e., penicillins/cephalosporins, carbapenems, aminoglycosides, and fluoroquinolones), that isolate was considered multidrug resistant (MDR). In accordance with the CLSI guidelines, P. aeruginosa ATCC 27853 and Escherichia coli ATCC 25922 were used as control strains in all susceptibility assays.
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