Meropenem (MEM) and ceftazidime (CAZ) were obtained from USP (Rockville, MD, USA). Penicillin G (PEN), cefoxitin (FOX), cefaclor (CEC), ceftriaxone (CTRX), cefepime (FEP) and tazobactam (TAZ) were from Solarbio (Beijing, China). Avibactam (AVI), vaborbactam, fosfomycin (FOS), amikacin (AMK) and imipenem (IMI) were purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA). The purity of all antibiotics used in this study was higher than 95%. The propylamino-derivatized controlled pore glass (CPG) beads, with a diameter of 125–140 µm and pore size of 50 nm, were obtained from Steinachglas (Steinach, Germany). HEPES was purchased from Sangon Biotech (Shanghai, China). All other chemical reagents were purchased from Sigma-Aldrich (Shanghai, China) unless stated otherwise. Molecular biology enzymes were bought from Takara Bio (Dalian, China), and the primers were purchased from Sangon Biotech (Shanghai, China).
Imipenem
Manufactured by MedChemExpress
Sourced in United States
Imipenem is a synthetic carbapenem antibiotic used as a laboratory standard for various applications. It functions as a reference compound for analytical and research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ceftriaxone, by Solarbio (1 mentions)
Penicillin g, by Solarbio (1 mentions)
Hepes, by Sangon (1 mentions)
Ceftazidime, by MedChemExpress (1 mentions)
Avibactam, by MedChemExpress (1 mentions)
Cefotaxime, by Sangon (1 mentions)
Carbenicillin, by Sangon (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Sangon (1 mentions)
Erythromycin, by Sangon (1 mentions)
Chloramphenicol, by Sangon (1 mentions)
4 protocols using imipenem
1
Antibiotic Susceptibility Testing Protocol
2
Multi-drug Resistant Pseudomonas Isolates Study
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Fifteen XDRPA isolates, defined as nonsusceptibility to ≥1 agent in all classes but ≤2 categories including colistin (18 (link)), collected from China-Japan Friendship Hospital from January 2017 to December 2020, were used in this study. The compounds of ceftazidime, avibactam, and imipenem were purchased from MedChemExpress (MCE). These drugs were dissolved in sterile solvents to generate a stock solution, and solutions with various concentrations were further prepared in Mueller-Hinton broth. Others (aztreonam, amikacin, meropenem, piperacillin-tazobactam) were from Wenzhou KONT Biology & Technology Co., Ltd.
3
Colorimetric Test for Carbapenemase Detection
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The Blue-Carba test was performed and interpreted as previously described (Pires et al., 2013 (link)). Briefly, 5 μL loopfuls of bacteria cultured on Mueller-Hinton agar (HuanKai, Guangzhou, China) were suspended in 0.04% bromothymol blue (Macklin, Shanghai, China) solution containing (test) or lacking (control) 3 mg/mL imipenem (MedChemExpress, New Jersey, United States) and 0.1 mM ZnSO4 (Damao, Tianjin, China). Color changes were registered after incubation at 37°C for 2 h. The result was considered positive when the solution containing imipenem became green or yellow and differed from the negative control. The result was negative if the solution lacking antibiotic presented the same or a stronger color change as the solution containing imipenem. A previously characterized NDM-5 producer (CQ02-121) was used as positive control (Sun et al., 2016 ) and a test tube containing only bacterial inoculum (E. coli ATCC 25922) and Blue-Carba solution was used as negative control for each isolate tested.
4
Bacterial Cultivation and Antibiotic Use
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This study complies with all relevant ethical regulations and the bacterial strains and plasmids used in this study are listed in Table S2 . Unless otherwise noted, S. aureus strains were grown in tryptic soy broth (TSB, Difco) at 37 °C with shaking (250 rpm), or on tryptic soy agar (TSA, Difco) at 37 °C. Escherichia coli strains were grown in Luria-Bertani (LB) broth at 37 °C with shaking (250 rpm) or on LB agar plates at 37 °C. For plasmid maintenance, antibiotics were used at the following concentrations where appropriate: for S. aureus, erythromycin (Sangon Biotech) at 10 μg/ml for RN4220 and 80 μg/ml for USA300 LAC and its derivatives, chloramphenicol (Sangon Biotech) at 15 μg/ml; for E. coli, carbenicillin (Sangon Biotech) at 150 μg/ml. For other reagents, tunicamycin and vancomycin (Dalian Meilun Biotech Co., Ltd.), targocil (Shanghai TopScience Co, Ltd.), oxacillin (Shanghai Aladdin Biochemical Technology Co.,Ltd.), imipenem, cefuroxime, ceftizoxime, cefaclor, cefoxitin, and epicatechin gallate from MedChemExpress, cefotaxime, isopropyl β-D-1-thiogalactopyranoside (IPTG), and AIP from Sangon Biotech, anhydrotetracycline (aTc) from APExBIO, Tarocin A1 from Merck, moenomycin complex from GlpBio, and nile red from Yeasen Biotechnology Co., Ltd. were used in this study.
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