J.Jpn. Surg. Soc.
ISSN 1880-1129

[Abstract] [Full Text PDF] （in Japanese / 2202KB） [Members Only And Two Factor Auth.]

J.Jpn. Surg. Soc.. 93(9): 914-917, 1992

Report on the annual meeting

RAPID DETECTION OF mecA GENE BY NESTED PCR FOR DIAGNOSIS OF METHICILIN RESISTANCE IN STAPHYLOCOCCUS AUREUS

1) Department of Surgery, Keio University, Tokyo, Japan
2) Laboratory Medicine, Keio University, Tokyo, Japan
3) Department of Microbiology, Meiji College of Pharmacy, Tokyo, Japan

Yuko Kitagawa1), Masakazu Ueda1), Nobutoshi Ando1), Kyuya Ishibiki1), Masaaki Kitajima1), Yoshio Kobayashi2), Toshihiko Arai3)

In recent years, the most common causative organism of hospital infections has been methicillin resistant Staphylococcus aureus (MRSA). The major mechanism of β-lactam resistance in MRSA is attributed to the production of a specific penicillin binding protein (PBP2'), which is a product of mecA gene, with extremely low binding affinities to β-lactams. In the present study, we have established a rapid identification method of MRSA by sensitive detection of mecA gene using nested PCR. Nested PCR method amplifying the target DNA in two steps enhanced the efficiency of the second round amplification. By means of this method, mecA gene was successfully detected in clinical samples, such as blood, pus, sputum and feces within 3-4 hrs. Rapid diagnosis of MRSA-bacteremia is paticularly important for prevention of sever systemic infection. There are some strains of S. aureus which possess mecA gene in spite of low minimal inhibitory concentration of DMPPC. In these strains expression of mecA gene is induced by contact of β-lactams and they obtain methicillin resistance. Using nested PCR method, these latent MRSA are rapidly and certainly detectable. This method should be useful for early and effective detection of MRSA hospital infections.

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