%0 Journal Article %A Cai, Lin %A Kong, Fanrong %A Wang, Qinning %A Wang, Huiping %A Xiao, Meng %A Sintchenko, Vitali %A Gilbert, Gwendolyn L. %T A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing %D 2009 %J Journal of Medical Microbiology, %V 58 %N 8 %P 1045-1057 %@ 1473-5644 %R https://doi.org/10.1099/jmm.0.007955-0 %K MRSA, meticillin-resistant S. aureus %K SCCmec, staphylococcal cassette chromosome mec %K mPCR/RLB, multiplex PCR-based reverse line-blot hybridization %K uPCR, uniplex PCR %K MLST, multilocus sequence typing %K MSSA, meticillin-sensitive S. aureus %I Microbiology Society, %X 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 Staphylococcus aureus and complete genome and partial SCCmec genes of seven non-MRSA strains, including meticillin-susceptible S. aureus and meticillin-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. %U https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.007955-0