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Rapid Molecular Typing of Methicillin-Resistant Staphylococcus aureus by PCR-RFLP

Published online by Cambridge University Press:  02 January 2015

Thomas A. Wichelhaus ,
Klaus-Peter Hunfeld ,
Boris Böddinghaus ,
Peter Kraiczy ,
Volker Schàfer  and
Volker Brade
Thomas A. Wichelhaus*
Affiliation:
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany
Klaus-Peter Hunfeld
Affiliation:
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany
Boris Böddinghaus
Affiliation:
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany
Peter Kraiczy
Affiliation:
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany
Volker Schàfer
Affiliation:
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany
Volker Brade
Affiliation:
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany
*
Institute of Medical Microbiology, University Hospital of Frankfurt, Paul-Ehrlich-Str. 40, 60596 Frankfurt am Main, Germany
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Abstract

Objective:

To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistant Staphylococcus aureus (MRSA) typing in routine epidemiological surveillance.

Design:

The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) following HaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent to mecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype.

Results:

Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates.

Conclusions:

This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability.

Type
Original Articles
Information
Infection Control & Hospital Epidemiology , Volume 22 , Issue 5 , May 2001 , pp. 294 - 298
Copyright
Copyright © The Society for Healthcare Epidemiology of America 2001

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