1887

Abstract

The aim of this study was to develop a new discriminatory method for MRSA SCC typing based on multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay to enable rapid identification and classification of MRSA SCC 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 to 14 whole-genome sequences and 18 partial SCC gene sequences of and complete genome and partial SCC genes of seven non-MRSA strains, including meticillin-susceptible 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 SCC types and 26 subtypes, including established and new subtypes of SCC III, IV (eight subtypes each) and V (three subtypes). The discriminatory power of mPCR/RLB SCC typing was similar to that of MLST and 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 SCC 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 SCC typing from passive epidemiological library typing into a potential tool for near-real-time infection control surveillance and tracking of MRSA transmission in hospitals.

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2009-08-01
2020-11-24
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