HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (Papers in Press[PDF]) All Versions of this Article: jmm.0.007955-0v1 58/8/1045    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Google Scholar Articles by Cai, L. Articles by Gilbert, G. L PubMed PubMed Citation Articles by Cai, L. Articles by Gilbert, G. L Agricola Articles by Cai, L. Articles by Gilbert, G. L

A New Multiplex PCR-Based Reverse Line Blot Hybridization (mPCR/RLB) Assay for Rapid Staphylococcal Cassette Chromosome mec (SCCmec) Typing

¹ Department of Dermatology, Peking University People's Hospital, Beijing 100044, P. R. China;

² Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Rese;

³ Department of Dermatology, Tianjin Medical University General Hospital, Tianjin, P. R. China;

⁴ Life Science College, Peking University, Beijing, P. R. China

⁵ E-mail: l.gilbert{at}usyd.edu.au

Received November 12, 2008
Accepted May 5, 2009

The aim of this study was to develop a new discriminatory method for MRSA SCCmec typing based on multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay to enable rapid identification and classification of MRSA SCCmec types, in a clinical laboratory. Forty five primer sets and 49 probes were designed and tested in uniplex PCR (uPCR) and mPCR/RLB. Probes were compared in silico to 14 whole genome sequences and 18 partial SCCmec gene sequences of S. aureus and complete genome and partial SCCmec gene of seven non-MRSA strains, including methicillin susceptible S. aureus and methicillin resistant coagulase negative staphylococci. The method was tested on a set of 42 well-characterized reference MRSA strains. It identified all five epidemiologically relevant SCCmec types and 26 subtypes, including established and new subtypes of SCCmec III, IV (eight subtypes each) and V (three subtypes). The discriminatory power of mPCR/RLB SCCmec typing was similar to that of MLST and spa typing (Simpson indices of diversity of 0.916, 0.926 and 0.882, respectively; differences not statistically significant). The application of mPCR/RLB hybridization assay to MRSA SCCmec typing can improve the specificity, discriminatory power and throughput of the typing procedure. The detection of up to 43 mPCR products in a single hybridization assay transforms MRSA SCCmec typing from passive epidemiological library typing into a potential tool for near real-time infection control surveillance and tracking of MRSA transmission in hospitals.