J Korean Soc Microbiol.  2000 Aug;35(4):289-297.

Comparison of infrequent restriction site-polymerase chain reaction and pulsed-field gel electrophoresis for molecular typing of Staphylococcus aureus and Escherichia coli

• Shin WS
• Kim TG
• Choi JH
• Lee DG
• Choi HB
• Yoo JH
• Kim JH
• Kang JH
• Min WS

Abstract

BACKGROUND
Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) are major pathogens in community and hospital. And they sometimes cause the outbreak in hospital in the immunocompromised patients. Pulsed-field gel electrophoresis (PFGE) has been regarded as a standard method for genotyping in epidemiologic studies, but it is laborious and time-consuming. Infrequent restriction site-polymerase chain reaction (IRS-PCR), a new genotyping methods, was performed to compare the applicability with PFGE. METHODS: We performed PFGE and IRS-PCR on S. aurues (n=120) and E. coli (n=117) which were collected clinically in 4 different hospitals. We assessed each method in terms of discriminatory power, quality, and efficiency. RESULTS: In E. coli, the discriminatory power of IRS-PCR was 46.7apprx86.7%, and that of PFGE was 88.9apprx96.7% according to hospital. But in S. aurues, the discriminatory power of IRS-PCR was 20apprx56.7%, and that of PFGE was 40apprx90% according to hospital. The typicality and reproducibility of IRS-PCR were 100% of each. PFGE needed four days to complete the procedure, but IRS-PCR could be performed within one day, IRS-PCR showed better resolution than PFGE. CONCLUSION: In case of gram negative bacteria (like E. coli), IRS-PCR could be a reliable alternative for epidemiologic typing due to better efficiency and comparable discriminatory power. But in the case of gram positive bacteria (like S. aureus), IRS-PCR does not seem to be suitable for the strain-to-strain differentiation. More trials and changes of restriction enzymes or primers could reveal the efficacy of IRS-PCR in the field of molecular typing.

MeSH Terms

Electrophoresis, Gel, Pulsed-Field*
Escherichia coli*
Escherichia*
Gram-Negative Bacteria
Gram-Positive Bacteria
Immunocompromised Host
Molecular Typing*
Staphylococcus aureus*
Staphylococcus*