Open AccessSmaI Restriction Site-Based Multiplex PCR for Typing of Hospital- and Community-Acquired Staphylococcus aureus
Author(s) -
Ibrahim Alzahrani,
Clare Hamson,
David Edge,
Jennifer Collins,
John D. Perry,
Muhammad W. Raza,
Kate Gould,
Colin R. Harwood
Publication year - 2011
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.00857-11
Subject(s) - multilocus sequence typing , typing , multiplex polymerase chain reaction , pulsed field gel electrophoresis , biology , multiplex , staphylococcus aureus , microbiology and biotechnology , methicillin resistant staphylococcus aureus , staphylococcal infections , sccmec , genetics , polymerase chain reaction , genotype , gene , bacteria
Methicillin-resistantStaphylococcus aureus (MRSA) is an important nosocomial pathogen, and morbidity and mortality rates associated with this pathogen have increased markedly in recent years. MRSA strains are generally resistant to several classes of antibiotics and are therefore difficult and costly to treat. A major issue is to identify the sources of MRSA infections and to monitor their epidemic spread. In this study, we report the development of a typing technique forS. aureus , based on single-nucleotide polymorphism (SNP) variations in and around SmaI-restriction sites (CCCGGG). An assessment of the SmaI restriction site-based multiplex PCR (SmaI-multiplex PCR) typing (SMT) with respect to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) revealed a high level of concordance in the clustering of the test strains. The SmaI-multiplex PCR was found to be more discriminatory than MLST/staphylococcal cassette chromosomemec (SCCmec ) typing but less discriminatory than PFGE. SMT can provide real-time information for the investigation of ongoingS. aureus hospital outbreaks. SMT meets the criteria of a practical typing method: it is simple, reproducible, and highly discriminatory and does not require expensive equipment or specialist expertise. Consequently, SmaI-multiplex PCR has the potential to be used in routine clinical microbiology laboratories.