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Multiplex PCR amplification assay for the detection of blaSHV, blaTEM and blaCTX-M genes in Enterobacteriaceae

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Abstract:

Monstein HJ, Östholm-Balkhed Å, Nilsson NV, Nilsson M, Dornbusch K, Nilsson LE. Multiplex PCR amplification assay for rapid detection of blaSHV, blaTEM and blaCTX-M genes in Enterobacteriaceae. APMIS 2007;115:1400–8.

Extended-spectrum β-lactamases (ESBLs) are often mediated by bla-SHV, blaTEM and blaCTX-M genes in Enterobacteriaceae and other Gram-negative bacteria. Numerous molecular typing methods, including PCR-based assays, have been developed for their identification. To reduce the number of PCR amplifications needed we have developed a multiplex PCR assay which detects and discriminates between bla-SHV, blaTEM and blaCTX-M PCR amplicons of 747, 445 and 593 bp, respectively. This multiplex PCR assay allowed the identification of bla-SHV, blaTEM and blaCTX-M genes in a series of clinical isolates of Enterobacteriaceae with previously characterised ESBL phenotype. The presence of blaSHV, blaTEM and blaCTX-M genes was confirmed by partial DNA sequence analysis. Apparently, the universal well-established CTX-M primer pair used here to reveal plasmid-encoded blaCTX-M genes would also amplify the chromosomally located K-1 enzyme gene in all Klebsiella oxytoca strains included in the study.

Keywords: ESBL; GenomiPhi plasmid DNA; automated DNA extraction; blaCTX-M genes; blaSHV; blaTEM genes; multiplex PCR assay

Document Type: Research Article

DOI: http://dx.doi.org/10.1111/j.1600-0463.2007.00722.x

Affiliations: Divisions of Clinical Microbiology

Publication date: December 1, 2007

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