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I restriction site-based multiplex PCR for typing of hospital and community acquired
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Dr John Perry
Dr Muhammad Raza
Professor Colin Harwood
Al-Zahrani IA, Hamson C, Edge D, Collins J, Perry JD, Raza M, Gould K, Harwood CR
Journal of Clinical Microbiology
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Methicillin resistant Staphylococcus 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 for S. aureus, based on single nucleotide polymorphism (SNP) variations in and around SmaI-restriction sites (CCCGGG). An assessment of the SmaI-multiplex PCR typing (SMT) with respect to Pulsed-Field Gel Electrophoresis (PFGE) and Multi-locus 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/SCCmec typing, but less discriminatory than PFGE. SMT can provide real-time information for the investigation of ongoing S. aureus hospital outbreaks. SMT meets the criteria of a practical typing method: it is simple, reproducible, 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.
American Society for Microbiology
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