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Journal of Clinical Microbiology, August 2002, p. 2786-2790, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2786-2790.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Methicillin-Resistant Staphylococci from Blood Culture Bottles by Using a Multiplex PCR Assay

L. Louie,1* J. Goodfellow,1 P. Mathieu,2 A. Glatt,2,3 M. Louie,1,4,{dagger} and A. E. Simor1,4

Sunnybrook and Women's College Health Sciences Centre,1 University of Toronto, Toronto, Ontario, Canada,4 St. Vincent Catholic Medical Centers, Brooklyn/Queens Region, Jamaica,2 New York Medical College, Valhalla, New York3

Received 7 February 2002/ Returned for modification 2 April 2002/ Accepted 15 May 2002

Rapid detection and accurate identification of methicillin-resistant staphylococci are critical for the effective management of infections caused by these organisms. We describe a multiplex PCR-based assay for the direct detection of methicillin-resistant staphylococci from blood culture bottles (BacT/Alert; Organon-Teknika, Durham, N.C.). A simple lysis method followed by a multiplex PCR assay designed to detect the nuc, mecA, and bacterial 16S rRNA genes was performed. A total of 306 blood culture specimens were collected over a period of 10 months from June 1998 to April 1999, consisting of 236 blood cultures growing staphylococci (including 124 methicillin-resistant Staphylococcus spp.), 50 positive blood cultures which grew organisms other than staphylococci, and 20 blood cultures that were negative for bacterial and fungal pathogens after 5 days of incubation and terminal subculture. DNA extraction, PCR, and detection could be completed in 2.5 h. Of the positive blood cultures with staphylococci, the multiplex PCR assay had a sensitivity and specificity of 99.2% and 100%, respectively. Our results show that rapid, direct detection of methicillin-resistant staphylococci is possible, allowing clinicians to make prompt and effective decisions for the management of patients with staphylococcal bacteremia.

* Corresponding author. Mailing address: Dept. of Microbiology, Sunnybrook and Women's College Health Sciences Centre, B121-2075 Bayview Ave., Toronto, Ontario, Canada M4N 3M5. Phone: (416) 480-4242. Fax: (416) 480-6845. E-mail: lisa.louie{at}swchsc.on.ca.

{dagger} Present address: Microbiology and Public Health, University of Alberta Hospital, Edmonton, Alberta T6G 2J2, Canada.

Journal of Clinical Microbiology, August 2002, p. 2786-2790, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2786-2790.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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