Bacterial identification was performed according to standard microbiological procedures. Antibiotic susceptibility was performed using the VITEK-2 Compact and the disc diffusion method. All microbiological methods were acted in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines of the corresponding year and the CLSI breakpoints of the corresponding year was used to identify the antimicrobial susceptibility. A 15-µg disc of tigecycline (Oxoid Ltd, Cambridge, UK) was used to determine susceptibility, and the breakpoints suggested by the FDA for Enterobacteriaceae (susceptible ≥19 mm; intermediate 15–18 mm, and resistant ≤14 mm), and gram-positive microorganisms (susceptible ≥19 mm) were used.18 (link) tigecycline clinical breakpoints using the disc-diffusion method against A. baumannii have not been established by both the CLSI and the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Our laboratory applied the clinical breakpoints recommended by the expert consensus of operating procedures for tigecycline in vitro sensitivity test published in 2013 (susceptible ≥16 mm, intermediate 13–15 mm, and resistant ≤12 mm).19 (link)
Tigecycline
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States, Belgium
Tigecycline is a broad-spectrum antibiotic used in the treatment of various bacterial infections. It functions as an inhibitor of bacterial protein synthesis, thereby preventing the growth and reproduction of pathogenic bacteria. Tigecycline is a semi-synthetic derivative of the tetracycline class of antibiotics.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by Thermo Fisher Scientific (19 mentions)
Gentamicin, by Thermo Fisher Scientific (15 mentions)
Trimethoprim sulfamethoxazole, by Thermo Fisher Scientific (14 mentions)
Imipenem, by Thermo Fisher Scientific (13 mentions)
Tetracycline, by Thermo Fisher Scientific (13 mentions)
Meropenem, by Thermo Fisher Scientific (12 mentions)
Ceftazidime, by Thermo Fisher Scientific (11 mentions)
Cefoxitin, by Thermo Fisher Scientific (11 mentions)
Amikacin, by Thermo Fisher Scientific (11 mentions)
Cefotaxime, by Thermo Fisher Scientific (10 mentions)
Cefepime, by Thermo Fisher Scientific (10 mentions)
Piperacillin tazobactam, by Thermo Fisher Scientific (10 mentions)
Ampicillin, by Thermo Fisher Scientific (9 mentions)
Linezolid, by Thermo Fisher Scientific (9 mentions)
Erythromycin, by Thermo Fisher Scientific (9 mentions)
Vancomycin, by Thermo Fisher Scientific (8 mentions)
Chloramphenicol, by Thermo Fisher Scientific (7 mentions)
Levofloxacin, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Ceftriaxone, by Thermo Fisher Scientific (5 mentions)
36 protocols using tigecycline
1
Antibiotic Susceptibility Testing of Bacteria
2
Antimicrobial Susceptibility Testing of Enterobacterales and S. aureus
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For all Enterobacterales and S. aureus, antimicrobial susceptibility testing was performed using Kirby-Bauer disk diffusion method on Mueller-Hinton agar according to the 2019 European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines (EUCAST: EUCAST, 2019 ). Enterobacterales were tested against ampicillin, cefotaxime, ceftazidime, meropenem, ciprofloxacin, gentamicin, tigecycline and sulfamethoxazole/trimethoprim (all Oxoid, Basingstoke, United Kingdom). ciprofloxacin resistance was defined as a MIC > 0.06 mg/L for S. enterica, confirmed by E-test (Oxoid, Basingstoke, United Kingdom) and a MIC > 1 mg/L for other Enterobacterales. A positive ESBL phenotype was confirmed by the double-disk diffusion test with cefotaxime and ceftazidime alone and in combination with clavulanic acid (Becton, Dickinson and Company, Sparks, MD, United States) as described before by the EUCAST (EUCAST, 2020 : Resistance mechanisms). S. aureus isolates were tested against penicillin, cefoxitin, clindamycin, erythromycin, ciprofloxacin, tetracycline, sulfamethoxazole/trimethoprim, tigecycline, gentamicin, rifampicin, linezolid, teicoplanin and vancomycin (all Oxoid, Basingstoke, United Kingdom).
3
Antibiotic Susceptibility Profiling of Clinically Relevant Pathogens
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Antibiotic susceptibility tests of clinically important bacteria and fungi were performed and interpreted according to the criteria of the Clinical and Laboratory Standards Institute [13 ] for the corresponding year, and the manufacturers’ instructions were followed for the use of antibiotics.
To assess the resistance of pathogens to other antibiotics, the K-B disk diffusion method was applied. Nine antibiotics were selected: piperacillin, cefoperazone/sulbactam, sulfamethoxazole, ceftazidime, cefotaxime, imipenem, gentamicin, ciprofloxacin, and tigecycline (Oxoid, UK). Polymyxin B was not included in antibiotic susceptibility tests because a preliminary test showed no resistance among the collected strains (Table S1).
To assess the resistance of pathogens to other antibiotics, the K-B disk diffusion method was applied. Nine antibiotics were selected: piperacillin, cefoperazone/sulbactam, sulfamethoxazole, ceftazidime, cefotaxime, imipenem, gentamicin, ciprofloxacin, and tigecycline (Oxoid, UK). Polymyxin B was not included in antibiotic susceptibility tests because a preliminary test showed no resistance among the collected strains (Table S1).
4
Antimicrobial Resistance Profiling
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Positive blood culture isolates were identified using VITEK 2 automated system (bioMerieux, Durham, North Carolina). MIC was confirmed using E-test (bioMerieux, France) in case of VRE. In case of any discrepancy E-test result was taken as final. Susceptibility testing was performed by Kirby Bauer method for tigecycline (15 μg, Oxoid Ltd., Basingstoke, Hampshire, England), and colistin (10 μg, HiMedia Laboratories, Mumbai) and results interpreted as per Clinical Laboratory Standards Institute (CLSI) guidelines2 and British society for Antimicrobial Chemotherapy (BSAC) guidelines where ever CLSI guidelines were not available.3 Daptomycin MIC was determined for Staphylococcus aureus and Enterococcus spp. using E-test during the period of 2010 and 2011. Antibiotic resistance data were extracted from the hospital information system (Intersystems, Cambridge, MA, USA) using a software Speedminer (Petaling Jaya, Malaysia) and analysed using a customized software, Wattal-Protech, Delhi, India.
5
Antimicrobial Resistance Profiling of Bacterial Isolates
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All strains were analysed for antimicrobial susceptibility with the Kirby-Bauer disk diffusion method on Mueller-Hinton agar including the antibiotics cefoxitin, clindamycin, erythromycin, penicillin, linezolid, ciprofloxacin, tetracycline, trimethoprim-sulfamethoxazole, tigecycline, gentamicin, and vancomycin (Oxoid, Basingstoke, United Kingdom) according to the 2015 European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines (www.eucast.org ). MRSA isolates were additionally tested for vancomycin resistance by E-test [17 (link)]. Isolates exhibiting resistance to three or more antimicrobial classes were defined as multidrug resistant (MDR), as described before [18 (link)].
6
Antimicrobial Susceptibility of A. baumannii
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To determine the antimicrobial susceptibility of A. baumannii isolates, the disc diffusion method was performed and interpreted according to the Clinical and Laboratory Standards Institute recommendations [17 ]. The following antibiotics were used in this study: amikacin (30 μg/disc), cefotaxime (30 μg/disc), ceftazidime (30 μg/disc), ceftriaxone (30 μg/disc), cefepime (30 μg/disc), ciprofloxacin (5 μg/disc), gentamicin (10 μg/disc), imipenem (10 μg/disc), meropenem (10 μg/disc), trimethoprim/sulfamethoxazole (1.25/23.75 μg/disc), tetracycline (30 μg/disc), cefoperazone/sulbactam (105 μg/disc), piperacillin/tazobactam (100/10 μg/disc), and tigecycline (15 μg/disc) (Oxoid, Basingstoke, UK). The presence of beta-lactamase encoding genes including blaOXA-23, blaOXA-24, blaOXA-58, and blaNDM-1 were detected using singlet PCR assay in all isolates of A. baumannii as described previously [16 (link)].
7
Antimicrobial Susceptibility Testing Protocol
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The antimicrobial susceptibility testing of the isolated bacterial species was determined by Kirby-Bauer disk diffusion method22 except against vancomycin and polymyxin B where antimicrobial susceptibility was determined by the broth microdilution method.23 The selection of the antimicrobial agents was based on the type of the organism being tested, and the results were interpreted following the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial susceptibility Testing (EUCAST) interpretive criteria.24 ,25 The antibiotic disks included in the study were gentamicin 10 µg, piperacillin/tazobactam 100 µg/10 µg, imipenem 10 µg, meropenem 10 µg, cefazolin 30 µg, ceftazidime 30 µg, cefepime 30 µg, cefoxitin 30 µg, ciprofloxacin 5 µg, sulfamethoxazole/trimethoprim 23.75 µg/1.25 µg, tigecycline 15 µg, aztreonam 30 µg, chloramphenicol 30 µg, doxycycline 30 µg, fusidic acid 10 µg, clindamycin 2 µg, erythromycin 15 µg, linezolid 30 µg and quinupristin/dalfopristin 15 µg (All from Oxoid Ltd., Hampshire, UK).
8
Antimicrobial Susceptibility of Bacterial Isolates
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Ten microliters of 7 isolates of bacterial extract (500 mg/ml) were added to a paper disc (6-mm diameter of Whatman no. 3) and air-dried. A sterile cotton swab was dipped in the antibiotic-resistant bacterial suspension on 0.85% normal saline solution and was streaked over the entire surface of the MHA medium, ensuring an even distribution of the inoculum. The antimicrobial discs (vancomycin, tigecycline, ampicillin, ceftazidime, and ceftazidime/clavulanic acid (Oxoid, England)) and DMSO were used as positive and negative controls, respectively. Then, they were incubated at 37°C for 24 h. After incubation, inhibition zone diameter was measured in millimetres using a ruler. Two independent experiments of disk diffusion assay were performed.
9
Antimicrobial Susceptibility of Enterococcus faecalis
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The antimicrobial susceptibility of the 76 E. faecalis isolates was tested by broth microdilution using 13 antimicrobials. Susceptibility to vancomycin (0.25–128 mg l−1), linezolid (0.25–128 mg l−1), penicillin (0.25–128 mg l−1), ampicillin (0.125–64 mg l−1), gentamicin (2–1024 mg l−1), streptomycin (4–2,048 mg l−1), kanamycin (4–2,048 mg l−1), tetracycline (0.25–128 mg l−1), tigecycline (0.03–2 mg l−1), erythromycin (0.125–64 mg l−1), lincomycin (0.5-256 mg l−1), ciprofloxacin (0.125-64 mg l−1), and chloramphenicol (0.25–128 mg l−1) (Oxoid, UK) was determined. E. faecalis ATCC 29212 was used as a quality control strain.
Susceptibility tests were performed according to recommendations by the Clinical and Laboratory Standards Institute (CLSI, 2015a ). MICs (minimal inhibitory concentrations) were evaluated based on the interpretative criteria of CLSI (CLSI, 2015c ) supplement VET01S for vancomycin, penicillin, ampicillin, erythromycin, tetracycline, doxycycline, and chloramphenicol for Enterococcus spp.; CLSI (CLSI, 2015b ) document M100-S25 for ciprofloxacin for Enterococcus spp.; and Comité de l´antibiogramme de la Société Française de Microbiologie (CA-SFM ) (CA-SFM, 2015 ) for lincomycin and all aminoglycosides for Streptococcus spp.
Susceptibility tests were performed according to recommendations by the Clinical and Laboratory Standards Institute (CLSI, 2015a ). MICs (minimal inhibitory concentrations) were evaluated based on the interpretative criteria of CLSI (CLSI, 2015c ) supplement VET01S for vancomycin, penicillin, ampicillin, erythromycin, tetracycline, doxycycline, and chloramphenicol for Enterococcus spp.; CLSI (CLSI, 2015b ) document M100-S25 for ciprofloxacin for Enterococcus spp.; and Comité de l´antibiogramme de la Société Française de Microbiologie (
10
Antibiotic Susceptibility of Enterobacteriaceae
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Coliforms were isolated from the floor and escalator transport samples by using CHROMagar ECC agar (France) in accordance with the manufacturer’s protocol. Taxonomic identification of the isolates was performed by sequencing the full-length 16S rRNA gene (Text S1 ). Antibiotic susceptibility to meropenem, cephalothin, tigecycline, and ertapenem (Oxoid, UK) was determined for the identified Enterobacteriaceae species using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST, version 10.0; https://www.eucast.org/ ) and the Clinical and Laboratory Standards Institute (CLSI 2015, M100-S25; https://clsi.org/ ). Reference strain Escherichia coli DH5α served as the quality control strain.
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