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Journal of Clinical Microbiology, January 2003, p. 254-260, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.254-260.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Methicillin-Resistant Staphylococcus aureus Directly from Sterile or Nonsterile Clinical Samples by a New Molecular Assay

Patrice Francois,1* Didier Pittet,2 Manuela Bento,1 Béatrice Pepey,3 Pierre Vaudaux,1 Daniel Lew,1,3 and Jacques Schrenzel1,3

Genomic Research Laboratory,1 Infection Control Program,2 Clinical Microbiology Laboratory, Division of Infectious Diseases, University Hospitals of Geneva, CH-1211 Geneva 14, Switzerland3

Received 15 April 2002/ Returned for modification 13 July 2002/ Accepted 19 October 2002

A rapid procedure was developed for detection and identification of methicillin-resistant Staphylococcus aureus (MRSA) directly from sterile sites or mixed flora samples (e.g., nose or inguinal swabs). After a rapid conditioning of samples, the method consists of two main steps: (i) immunomagnetic enrichment in S. aureus and (ii) amplification-detection profile on DNA extracts using multiplex quantitative PCR (5'-exonuclease qPCR, TaqMan). The triplex qPCR assay measures simultaneously the following targets: (i) mecA gene, conferring methicillin resistance, common to both S. aureus and Staphylococcus epidermidis; (ii) femA gene from S. aureus; and (iii) femA gene from S. epidermidis. This quantitative approach allows discrimination of the origin of the measured mecA signal. qPCR data were calibrated using two reference strains (MRSA and methicillin-resistant S. epidermidis) processed in parallel to clinical samples. This 96-well format assay allowed analysis of 30 swab samples per run and detection of the presence of MRSA with exquisite sensitivity compared to optimal culture-based techniques. The complete protocol may provide results in less than 6 h (while standard procedure needs 2 to 3 days), thus allowing prompt and cost-effective implementation of contact precautions.

* Corresponding author. Mailing address: Division of Infectious Diseases/Genomic Research Laboratory, University Hospitals of Geneva, CH-1211 Geneva 14, Switzerland. Phone: (41)-22 372 9338. Fax: (41)-22 372 9830. E-mail: patrice.francois{at}hcuge.ch.

Journal of Clinical Microbiology, January 2003, p. 254-260, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.254-260.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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