title: SmaI restriction site-based multiplex PCR for typing of hospital- and community-acquired Staphylococcus aureus.
abstract: 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 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 chromosome mec (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, 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.
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Medical School, Newcastle University, Newcastle upon Tyne NE2 4AX, UK.
authors.forename: Ibrahim A
authors.initials: IA
authors.lastname: Al-Zahrani
authors.forename: Clare
authors.initials: C
authors.lastname: Hamson
authors.forename: David
authors.initials: D
authors.lastname: Edge
authors.forename: Jennifer
authors.initials: J
authors.lastname: Collins
authors.forename: John D
authors.initials: JD
authors.lastname: Perry
authors.forename: Muhammad
authors.initials: M
authors.lastname: Raza
authors.forename: Kate
authors.initials: K
authors.lastname: Gould
authors.forename: Colin R
authors.initials: CR
authors.lastname: Harwood
mesh_terms: D016000:Cluster Analysis; D017714:Community-Acquired Infections / Q000382:microbiology; D003428:Cross Infection / Q000382:microbiology; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D005838:Genotype; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D058885:Multilocus Sequence Typing / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D013203:Staphylococcal Infections / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification