Antimicrobial susceptibility testing of CR-HMKP isolates was initially performed using the VITEK 2 compact system. Subsequently, ampicillin, amikacin, aztreonam, cefazolin, cefepime, ceftazidime, ceftriaxone, ciprofloxacin, levofloxacin, gentamicin, trimethoprim-sulfamethoxazole, tobramycin (National Institutes for Food and Drug Control, Beijing, China); ertapenem, imipenem (Merck Sharp & Dohme Corp, Hangzhou, China); meropenem (Sumitomo Pharmaceuticals, Suzhou, China); and piperacillin-tazobactam (Pfizer, New York, NY, USA). MICs were confirmed using the agar dilution method according to the recommendations and breakpoints proposed by the CLSI [13 14 ]. Tigecycline (Pfizer) and colistin (Sigma Aldrich, Shanghai, China) MICs were determined by the broth microdilution method, using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines and breakpoint [15 ]. The standard strains Escherichia coli ATCC25922 and Pseudomonas aeruginosa ATCC27853 were used for quality control.
Piperacillin tazobactam
Manufactured by Pfizer
Sourced in China
Piperacillin/tazobactam is a combination of two antimicrobial agents, piperacillin and tazobactam. It is a broad-spectrum antibiotic used to treat a variety of bacterial infections. The core function of piperacillin/tazobactam is to inhibit bacterial cell wall synthesis, thereby preventing the growth and proliferation of susceptible bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Tigecycline, by Pfizer (4 mentions)
Imipenem, by Merck Group (3 mentions)
Ciprofloxacin, by Bayer (3 mentions)
Ceftriaxone, by Roche (3 mentions)
Colistin, by Merck Group (1 mentions)
Ertapenem, by Merck Group (1 mentions)
Polymyxin b, by Avantor (1 mentions)
Maldi tof ms, by Bruker (1 mentions)
Cefoperazone sulbactam, by Pfizer (1 mentions)
Imipenem, by MSD (1 mentions)
5 protocols using piperacillin tazobactam
1
Antimicrobial Susceptibility Testing of CR-HMKP
2
Antimicrobial Susceptibility Testing Protocol
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All isolates were identified with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) (Bruker Daltonics, Bremen, Germany). Minimum inhibitory concentrations (MICs) were determined by the agar dilution method according to CLSI guidelines (M100-S27). The tested drugs included ceftriaxone (Roche China, Shanghai, China), cefotaxime, ceftazidime, cefepime, aztreonam, amikacin, levofloxacin, minocycline, fosfomycin (National Institute for Food and Drug Control of China, Beijing, China), piperacillin/tazobactam, tigecycline (Pfizer, NY, USA), imipenem (Merck Sharp & Dohme, Hangzhou, China), meropenem (Sumitomo Pharmaceuticals, Suzhou, China), ciprofloxacin (Bayer, Leverkusen, Germany), and polymyxin B (Amresco, Solon, USA). Strains used in quality control were Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853. The results were interpreted according to 2017 CLSI standards (M100-S27). The tigecycline test was performed according to the Food and Drug Administration standards.
3
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility was determined by the agar dilution method and broth microdilution method (only for tigecycline), and the results were interpreted according to the minimum inhibitory concentration (MIC) interpretive breakpoints recommended by the Clinical and Laboratory Standards Institute (CLSI) in 2017.10 The tested antimicrobial agents included amikacin, ampicillin, ceftazidime, chloramphenicol, cefotaxime, erythromycin, cefepime, cefoxitin, levofloxacin, moxifloxacin, minocycline, rifampicin, sulfamethoxazole, teicoplanin, vancomycin (National Institute for Food and Drug Control of China, Beijing, China), ciprofloxacin (Bayer AG, Leverkusen, Germany), ceftriaxone (Hoffman-La Roche Ltd., Basel, Switzerland), cefoperazone/sulbactam, piper-acillin/tazobactam, tigecycline (Pfizer, Inc., New York, NY, USA), imipenem (Merck & Co., Inc., Whitehouse Station, NJ, USA), and meropenem (Sumitomo Dainippon Pharma, Osaka, Japan). The tigecycline test was performed according to the Food and Drug Administration standards. The double-disk synergy test was used to determine extended spectrum β-lactamase (ESBL) production among isolates of Escherichia coli and Klebsiella pneumoniae as recommended by the CLSI.10 The reference isolates E. faecalis ATCC 29212, S. aureus ATCC29213, E. coli ATCC 25922 and P. aeruginosa ATCC 27853 were used as quality control isolates.
4
Kidney Capsule Transplantation of Organoids
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As previously described, a single HIO, matured in vitro for 28 d, was removed from Matrigel and then transplanted under the kidney capsule12 (link). Briefly, the mice were anesthetized with 2% inhaled isoflurane (Butler Schein) and 2.5–3 l min−1 oxygen. The left side of the mouse was then prepped in sterile fashion with isopropyl alcohol and providine-iodine. A small left-posterior subcostal incision was made to expose the kidney. A subcapsular pocket was created in the kidney capsule and the HIO was then placed into the pocket. The kidney was then returned to the peritoneal cavity and the mice were given an intraperitoneal flush with 2–3 ml of piperacillin/tazobactam (100 mg kg−1; Pfizer) to help prevent bacterial infection. The skin was closed in a double layer. For pain control, mice were then given a subcutaneous injection with buprenorphine (0.05 mg kg−1; Midwest Veterinary Supply) or carprofen (4 mg kg−1; Midwest Veterinary Supply) and were monitored for the next 48 h following surgery. Additional injections of pain medication were given if needed. At 12 and 16 weeks following engraftment, the mice were then euthanized and the tissues were collected and analyzed.
5
Antimicrobial Resistance Profiling of E. coli
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The minimum inhibitory concentrations (MICs) of the following antimicrobials, including ceftriaxone (Roche, Shanghai, China), ceftazidime, cefoxitin, cefepime, amikacin (National Institutes for Food and Drug Control, Beijing, China), piperacillin–tazobactam, tigecycline (Pfizer, NY, USA), imipenem (MSD, Hangzhou, China), meropenem (Sumitomo Pharmaceuticals, Suzhou, China), ciprofloxacin (Bayer, Leverkusen, Germany) and colistin (Amresco, Solon, USA) were determined by agar dilution method, according to Clinical and Laboratory Standards Institute (CLSI) guidelines (Clinical and Laboratory Standards Institute, 2015 ). The results were interpreted according to CLSI breakpoints (Clinical and Laboratory Standards Institute, 2016 ). Polymerase chain reaction (PCR) was used to detect the presence of mcr-1, carbapenemase genes and other resistance genes as previously described (Wang et al., 2014 (link); Liu, Y. Y. et al., 2016 (link)). Multilocus sequence typing (MLST) was performed as described on the E. coli MLST website (http://mlst.warwick.ac.uk/mlst/dbs/Ecoli ).
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