Diagnostic Microbiology & Infectious Disease
Volume 45, Issue 1 , Pages 81-84, January 2003

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

  • Silvano Palladino

      Affiliations

    • 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.
    • ,
  • Ian D. Kay

      Affiliations

    • Department of Microbiology & Infectious Diseases, Royal Perth Hospital, Perth, Western Australia
    • ,
  • Anna Maria Costa

      Affiliations

    • Department of Microbiology & Infectious Diseases, Royal Perth Hospital, Perth, Western Australia
    • ,
  • Erica J. Lambert

      Affiliations

    • Department of Microbiology & Infectious Diseases, Royal Perth Hospital, Perth, Western Australia
    • ,
  • James P. Flexman

      Affiliations

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

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.

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PII: S0732-8893(02)00505-9

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

Diagnostic Microbiology & Infectious Disease
Volume 45, Issue 1 , Pages 81-84, January 2003

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