1887

Abstract

A multiplex TaqMan real-time PCR to detect carbapenem-hydrolysing class D β-lactamases ( -like, -like, -like and -like genes) was developed and evaluated for early detection of imipenem (IMP) resistance in clinically significant isolates. Well-characterized strains of were used as positive controls and non- strains were used to assess specificity. Analytical sensitivity was quantified by comparison with the number of bacterial c.f.u. Forty of 46 (87 %) clinically significant and IMP-resistant isolates were positive for the -like gene, and one isolate (2 %) was positive for the -like gene. The -like gene was not detected in any of the 46 IMP-resistant strains and the -like gene was identified in both IMP-resistant and non-resistant . All 11 non- bacteria produced a negative result for each of the four genes. This assay was able to detect as few as 10 c.f.u. per assay. This real-time PCR method demonstrated rapid detection of OXA-like carbapenem resistance in in comparison with phenotypic susceptibility testing methodology. This method could be adapted to a multiplexed single reaction for rapid detection of genes associated with carbapenem resistance in and potentially other clinically significant multidrug-resistant Gram-negative bacteria.

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2012-11-01
2020-01-21
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