Simple and reliable multiplex PCR assay for detection of blaTEM, blaSHV and blaOXA-1 genes in Enterobacteriaceae

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Abstract

Third-generation cephalosporin resistance is often mediated by TEM- and SHV-type β-lactamases in Enterobacteriaceae. TEM-type and OXA-1 enzymes are the major plasmid-borne β-lactamases implicated in amoxicillin-clavulanic acid resistance in Escherichia coli isolates. We have developed a rapid and simple multiplex polymerase chain reaction (PCR) which discriminates blaTEM, blaSHV and blaOXA-1 genes by generating fragments of 516, 392 and 619 bp respectively. Multiplex PCR analysis of 51 amoxicillin-clavulanate resistant E. coli isolates detected blaTEM and blaSHV genes in 45 and two strains, respectively, and only one strain harboured a blaOXA-1 gene. Twenty-three of the 40 cefotaxime-resistant Enterobacteriaceae isolates produced amplicons with a size compatible with the presence of blaTEM (13 strains), blaSHV (six strains) genes or the association of both genes (four strains). These results were verified by colony hybridisation. Therefore, multiplex PCR is a suitable tool for initial rapid screening of bla genes in Enterobacteriaceae. © 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

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Colom, K., Pérez, J., Alonso, R., Fernández-Aranguiz, A., Lariño, E., & Cisterna, R. (2003). Simple and reliable multiplex PCR assay for detection of blaTEM, blaSHV and blaOXA-1 genes in Enterobacteriaceae. FEMS Microbiology Letters, 223(2), 147–151. https://doi.org/10.1016/S0378-1097(03)00306-9

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