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doi: 10.1093/jac/dkaa031
pmid: 32073602
handle: 10261/228761 , 2445/176218 , 10067/1721250151162165141 , 10668/15128
doi: 10.1093/jac/dkaa031
pmid: 32073602
handle: 10261/228761 , 2445/176218 , 10067/1721250151162165141 , 10668/15128
Abstract Objectives To evaluate and compare the efficacy of real-time PCR (Xpert Carba-R) and loop-mediated isothermal amplification (Eazyplex® SuperBug CRE) for detecting carbapenemase carriage in Enterobacteriaceae directly from bronchoalveolar lavage (BAL). Methods Negative BAL samples were spiked with 21 well-characterized carbapenemase-producing Enterobacteriaceae strains to a final concentration of 102–104 cfu/mL. Xpert Carba-R (Cepheid, Sunnyvale, CA, USA), which detects five targets (blaKPC, blaNDM, blaVIM, blaOXA-48 and blaIMP-1), and the Eazyplex® SuperBug CRE system (Amplex-Diagnostics GmbH, Germany), which detects seven genes (blaKPC, blaNDM, blaVIM, blaOXA-48, blaOXA-181, blaCTXM-1 and blaCTXM-9), were evaluated for the detection of these genes directly from BAL samples. Results Xpert Carba-R showed 100% agreement with carbapenemase characterization by PCR and sequencing for all final bacteria concentrations. Eazyplex® SuperBug CRE showed 100%, 80% and 27% agreement with PCR and sequencing when testing 104, 103 and 102 cfu/mL, respectively. False negative results for Eazyplex® SuperBug CRE matched the highest cycle threshold values for Xpert Carba-R. Hands-on time for both assays was about 15 min, but Eazyplex® SuperBug CRE results were available within 30 min, whereas Xpert Carba-R took around 50 min. Conclusions We here describe the successful use of two commercial diagnostic tests, Xpert Carba-R and Eazyplex® SuperBug CRE, to detect bacterial carbapenem resistance genes directly in lower respiratory tract samples. Our results could be used as proof-of-concept data for validation of these tests for this indication.
Bacteria, Pharmacology. Therapy, Respiratory organs, Real-Time Polymerase Chain Reaction, Aparell respiratori, Bacteris, Sensitivity and Specificity, Diagnòstic microbiològic, beta-Lactamases, Bacterial Proteins, Enterobacteriaceae, Molecular Diagnostic Techniques, Germany, Diagnostic microbiology, Biology, Bronchoalveolar Lavage Fluid, Nucleic Acid Amplification Techniques
Bacteria, Pharmacology. Therapy, Respiratory organs, Real-Time Polymerase Chain Reaction, Aparell respiratori, Bacteris, Sensitivity and Specificity, Diagnòstic microbiològic, beta-Lactamases, Bacterial Proteins, Enterobacteriaceae, Molecular Diagnostic Techniques, Germany, Diagnostic microbiology, Biology, Bronchoalveolar Lavage Fluid, Nucleic Acid Amplification Techniques
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