A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB)   assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing

doi:10.1099/jmm.0.007955-0

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	A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing
	Cai, Lin ; Kong, Fanrong ; Wang, Qinning ; Wang, Huiping ; Xiao, Meng ; Sintchenko, Vitali ; Gilbert, Gwendolyn L.
刊名	journal of medical microbiology
	2009
关键词	AUREUS STRAINS METHICILLIN-RESISTANCE GENETIC DIVERSITY IDENTIFICATION LOCUS ASSIGNMENT AUSTRALIA VARIANTS STRATEGY ELEMENT
DOI	10.1099/jmm.0.007955-0
英文摘要	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.; Microbiology; SCI(E); PubMed; 9; ARTICLE; 8; 1045-1057; 58
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/307516]
专题	生命科学学院
推荐引用方式 GB/T 7714	Cai, Lin,Kong, Fanrong,Wang, Qinning,et al. A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing[J]. journal of medical microbiology,2009.
APA	Cai, Lin.,Kong, Fanrong.,Wang, Qinning.,Wang, Huiping.,Xiao, Meng.,...&Gilbert, Gwendolyn L..(2009).A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing.journal of medical microbiology.
MLA	Cai, Lin,et al."A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing".journal of medical microbiology (2009).