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. 2004 Dec;42(12):5739-44.
doi: 10.1128/JCM.42.12.5739-5744.2004.

Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets

H Sakai  1 , G W Procop, N Kobayashi, D Togawa, D A Wilson, L Borden, V Krebs, T W Bauer
Affiliations

Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets

H Sakai et al. J Clin Microbiol. 2004 Dec.

Abstract

A real-time PCR assay that uses two fluorescence resonance energy transfer probe sets and targets the tuf gene of staphylococci is described here. One probe set detects the Staphylococcus genus, whereas the other probe set is specific for Staphylococcus aureus. One hundred thirty-eight cultured isolates, which contained 41 isolates of staphylococci representing at least nine species, and 100 positive blood cultures that contained gram-positive cocci in clusters were tested. This assay was 100% sensitive and 100% specific for the detection of the Staphylococcus genus and of S. aureus.

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Figures

FIG. 1.
FIG. 1.
(a) Amplification curves with broad-range Staphylococcus FRET probes. The broad-range Staphylococcus FRET probes generated positive quantification curves for all of the staphylococci tested. Three strains of Enterococcus were distinctly negative in this study. (b) Melt peak analysis of broad-range Staphylococcus assay. Enterococcus faecium was not detected in the quantification mode while the melting curve was generated by melt peak analysis. The Tms of the staphylococci were all >58°C. The Tm of E. faecium was distinctively lower than those of the other CoNS.
FIG. 2.
FIG. 2.
Melt curves of S. aureus-specific assay. The melt curves were generated from S. aureus samples only.
See this image and copyright information in PMC

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