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Epidemiological characterization of methicillin-resistant Staphylococcus aureus isolated in the North West of England by protein A (spa) and coagulase (coa) gene polymorphisms

Published online by Cambridge University Press:  01 December 1998

J. WALKER
Affiliation:
Manchester Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester M20 2LR
R. BORROW
Affiliation:
Manchester Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester M20 2LR
V. EDWARDS-JONES
Affiliation:
Manchester Metropolitan University, Department of Biological Sciences, Faculty of Science and Engineering, Chester Street, Manchester M1 5GD
B. A. OPPENHEIM
Affiliation:
Manchester Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester M20 2LR
A. J. FOX
Affiliation:
Manchester Public Health Laboratory, Withington Hospital, Nell Lane, West Didsbury, Manchester M20 2LR
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Abstract

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In a comparative study, isolates of methicillin-resistant Staphylococcus aureus (MRSA) with known pulsed-field gel electrophoresis (PFGE) and bacteriophage type were analysed by polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLP) for additional discriminatory subtyping information. PFGE was previously performed using standardized, commercially available kits and pre-programmed software. Isolates were examined for coagulase (coa) and protein A (spa) gene polymorphisms following PCR amplification of the coa hypervariable and spa repeat regions. Coa gene RFLPs produced a total of 38 distinct combined patterns after digestion with HaeIII and AluI and identified the predominant epidemic (EMRSA) types 15 and 16. A unique HaeIII restriction site was identified by RFLP and sequence analysis in the coa gene for EMRSA 15 but not EMRSA 16. The spa gene PCR yielded a total of 14 different profiles ranging from 3–18 repeats with the 2 predominant EMRSA types falling into 2 distinct groups. PCR detection of coa and spa polymorphisms offer a rapid preliminary strain identification and discriminatory subtyping information for surveillance of MRSA.

Type
Research Article
Copyright
© 1998 Cambridge University Press