Rapid molecular typing of methicillin-resistant Staphylococcus aureus by PCR-RFLP.
OBJECTIVE
To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistant Staphylococcus aureus (MRSA) typing in routine epidemiological surveillance.


METHODS
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.
Wichelhaus
T A
TA
Institute of Medical Microbiology, University Hospital of Frankfurt, Frankfurt am Main, Germany.
Hunfeld
K P
KP
Böddinghaus
B
B
Kraiczy
P
P
Schäfer
V
V
Brade
V
V
D015373:Bacterial Typing Techniques; D003030:Coagulase / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods; D004269:DNA, Bacterial / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D005838:Genotype; D006801:Humans; D016106:Methicillin Resistance; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D013203:Staphylococcal Infections / Q000382:microbiology*; D013210:Staphylococcus / Q000145:classification* / Q000201:enzymology / Q000235:genetics
D003030:Coagulase; D004269:DNA, Bacterial