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Journal of Clinical Microbiology, October 2005, p. 5026-5033, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5026-5033.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Novel Multiplex PCR Assay for Characterization and Concomitant Subtyping of Staphylococcal Cassette Chromosome mec Types I to V in Methicillin-Resistant Staphylococcus aureus

Kunyan Zhang,1,2,3,4,5* Jo-Ann McClure,1 Sameer Elsayed,2,3,5 Thomas Louie,3,4 and John M. Conly1,2,3,4,5

Centre for Antimicrobial Resistance, Calgary Health Region/Calgary Laboratory Services/University of Calgary, Calgary, Alberta, Canada,1 Departments of Pathology & Laboratory Medicine,2 Microbiology and Infectious Diseases,3 Medicine, University of Calgary, Calgary, Alberta, Canada,4 Calgary Laboratory Services, Calgary, Alberta, Canada5

Received 21 April 2005/ Returned for modification 15 June 2005/ Accepted 15 July 2005

Staphylococcal cassette chromosome mec (SCCmec) typing is essential for understanding the molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA). SCCmec elements are currently classified into types I to V based on the nature of the mec and ccr gene complexes, and are further classified into subtypes according to their junkyard region DNA segments. Previously described traditional SCCmec PCR typing schemes require multiple primer sets and PCR experiments, while a previously published multiplex PCR assay is limited in its ability to detect recently discovered types and subtypes such as SCCmec type V and subtypes IVa, b, c, and d. We designed new sets of SCCmec type- and subtype-unique and specific primers and developed a novel multiplex PCR assay allowing for concomitant detection of the methicillin resistance (mecA gene) (also serving as an internal control) to facilitate detection and classification of all currently described SCCmec types and subtypes I, II, III, IVa, b, c, d, and V. Our assay demonstrated 100% sensitivity and specificity in accurately characterizing 54 MRSA strains belonging to the various known SCCmec types and subtypes, when compared with previously described typing methods. Further application of our assay in 453 randomly selected local clinical isolates confirmed its feasibility and practicality. This novel assay offers a rapid, simple, and feasible method for SCCmec typing of MRSA, and may serve as a useful tool for clinicians and epidemiologists in their efforts to prevent and control infections caused by this organism.

* Corresponding author. Mailing address: Department of Pathology & Laboratory Medicine, #9-3535 Research Road N.W., Calgary, Alberta, Canada T2L 2K8. Phone: 403-770-3583. Fax: 403-770-3347. E-mail: kunyan.zhang{at}cls.ab.ca.

Journal of Clinical Microbiology, October 2005, p. 5026-5033, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5026-5033.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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Copyright © 2005 by the American Society for Microbiology. All rights reserved.