Comparison of DNA fingerprinting by PFGE and PCR-RFLP of the coagulase gene to distinguish MRSA isolates.
Staphylococcus aureus isolates were collected from epidemiologically unrelated clinical sources in Japan between 1991 and 1993. A total of 40 isolates, five each of eight coagulase types, were analysed by polymerase chain reaction (PCR) of the coagulase gene, PCR-restriction fragment length polymorphism (RFLP) after AluI digestion, and pulsed-field gel electrophoresis (PFGE) of chromosomal DNA after SmaI digestion. The efficiency of discrimination among the isolates increased in the order of PCR < PCR-RFLP < PFGE, yielding five, 13 and 31 different types, respectively. To assess the clinical use of these methods, 42 additional methicillin-resistant S. aureus (MRSA) isolates collected from 27 inpatients in a hospital were analysed. PFGE and PCR-RFLP were able to discriminate 11 and four types, respectively. PFGE analysis detected cross-infection between four postoperative patients in an intensive-care unit, and in six neonates in intensive care. We conclude that of the three methods tested, PFGE analysis currently allows the most effective discrimination of MRSA strains.
Nada
T
T
Department of Clinical Laboratory Medicine, Nagoya University Hospital, Japan.
Ichiyama
S
S
Osada
Y
Y
Ohta
M
M
Shimokata
K
K
Kato
N
N
Nakashima
N
N
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D003030:Coagulase / Q000235:genetics*; D003428:Cross Infection / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis*; D016002:Discriminant Analysis; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D006801:Humans; D016106:Methicillin Resistance; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification* / Q000201:enzymology / Q000235:genetics
D003030:Coagulase; D004269:DNA, Bacterial