New Real-Time PCR Assay for Rapid Detection of Methicillin ...

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JOURNAL OF CLINICAL MICROBIOLOGY, May 2004, p. 1875–1884 0095-1137/04/$08.00⫹0 DOI: 10.1128/JCM.42.5.1875–1884.2004 Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Vol. 42, No. 5

New Real-Time PCR Assay for Rapid Detection of MethicillinResistant Staphylococcus aureus Directly from Specimens Containing a Mixture of Staphylococci A. Huletsky,1,2 R. Giroux,1 V. Rossbach,1 M. Gagnon,1 M. Vaillancourt,1 M. Bernier,1 F. Gagnon,1 K. Truchon,3 M. Bastien,1 F. J. Picard,1 A. van Belkum,4 M. Ouellette,1,2 P. H. Roy,1,5 and M. G. Bergeron1,2* Centre de Recherche en Infectiologie de l’Universite´ Laval, CHUQ,1 Division de Microbiologie, Faculte´ de Me´decine,2 and De´partement de Biochimie et de Microbiologie, Faculte´ des Sciences et de Ge´nie,5 Universite´ Laval, Sainte-Foy, Que´bec, Canada G1V 4G2; Infectio Diagnostic (I.D.I.) Inc., Sainte-Foy, Que´bec, Canada G1V 2K83; and Department of Medical Microbiology and Infectious Diseases, Erasmus MC, 3015 GD Rotterdam, The Netherlands4 Received 16 July 2003/Returned for modification 8 October 2003/Accepted 10 February 2004

Molecular methods for the rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) are generally based on the detection of an S. aureus-specific gene target and the mecA gene. However, such methods cannot be applied for the direct detection of MRSA from nonsterile specimens such as nasal samples without the previous isolation, capture, or enrichment of MRSA because these samples often contain both coagulasenegative staphylococci (CoNS) and S. aureus, either of which can carry mecA. In this study, we describe a real-time multiplex PCR assay which allows the detection of MRSA directly from clinical specimens containing a mixture of staphylococci in