Rapid Molecular Typing of Methicillin-ResistantStaphylococcus aureusby PCR-RFLP

Abstract

AbstractObjective:To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistantStaphylococcus aureus(MRSA) typing in routine epidemiological surveillance.Design:The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) followingHaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent tomecA. 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.