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Volume 45, Issue 1, Pages 81-84 (January 2003)

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Real-time PCR for the rapid detection of vanA and vanB genes

Silvano PalladinoaCorresponding Author Informationemail address, Ian D. Kaya, Anna Maria Costaa, Erica J. Lamberta, James P. Flexmana

Received 10 May 2002; accepted 26 August 2002.

Abstract 

A real-time PCR assay suitable for use on the Roche LightCycler platform was developed to replace an existing gel-based PCR assay for the simultaneous detection of the vanA & vanB genes in enterococcal isolates. Novel Fluorescence Resonance Energy Transfer (FRET) hybridization probes were designed. The multiplex real-time PCR assay and the existing gel-based assay were 100% concordant and both correctly detected the vanA or vanB genes in 4/4 VanA E. faecium and 25/25 VanB E. faecium. Additionally, 1/1 VanC1 E. gallinarum, 1/1 VanC2 E. casseliflavus and 47/47 vancomycin susceptible enterococci were negative for the vanA and vanB genes in both PCR assays. Results were available within 1.5 h for the real-time PCR assay compared to up to 5.5 h for the conventional PCR assay.

a Department of Microbiology & Infectious Diseases, Royal Perth Hospital, Perth, Western Australia

Corresponding Author InformationCorresponding author. Tel.: +61 8 9224 2444; fax: +61 8 9224 1799.

PII: S0732-8893(02)00505-9

doi:10.1016/S0732-8893(02)00505-9

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