Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae

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ID: 292002
2010
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Abstract
To develop a rapid and reliable tool to detect by multiplex PCR assays the most frequently widespread β-lactamase genes encoding the OXA-1-like broad-spectrum β-lactamases, extended-spectrum β-lactamases (ESBLs), plasmid-mediated AmpC β-lactamases and class A, B and D carbapenemases. Following the design of a specific group of primers and optimization using control strains, a set of six multiplex PCRs and one simplex PCR was created. An evaluation of the set was performed using a collection of 31 Enterobacteriaceae strains isolated from clinical specimens showing a resistance phenotype towards broad-spectrum cephalosporins and/or cephamycins and/or carbapenems. Direct sequencing from PCR products was subsequently carried out to identify β-lactamase genes. Under optimized conditions, all positive controls confirmed the specificity of group-specific PCR primers. Except for the detection of carbapenemase genes, multiplex and simplex PCR assays were carried out using the same PCR conditions, allowing assays to be performed in a single run. Out of 31 isolates selected, 22 strains produced an ESBL, mostly CTX-M-15 but also CTX-M-1 and CTX-M-9, SHV-12, SHV-5, SHV-2, TEM-21, TEM-52 and a VEB-type ESBL, 6 strains produced a plasmid-mediated AmpC β-lactamase (five DHA-1 and one CMY-2) and 3 strains produced both an ESBL (two SHV-12, one CTX-M-15) and a plasmid-mediated AmpC β-lactamase (DHA-1). We report here the development of a useful method composed of a set of six multiplex PCRs and one simplex PCR for the rapid screening of the most frequently encountered β-lactamases. This method allowed direct sequencing from the PCR products.
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openalex_W2141888371 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Caroline Dallenne, Anaelle Da Costa, Dominique Decré, Christine F. Favier, Guillaume Arlet
Journal The Journal of antimicrobial chemotherapy
Year 2010
DOI
10.1093/jac/dkp498
URL
Keywords Keywords not found

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