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Journal of Clinical Microbiology, December 2004, p. 5462-5466, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5462-5466.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a Rapid PCR Method Using the Insertion Sequence IS1203 for Genotyping Shiga Toxin-Producing Escherichia coli O157

Masahiro Suzuki,* Masakado Matsumoto, Mami Hata, Masao Takahashi, and Kenji Sakae

Department of Microbiology, Aichi Prefectural Institute of Public Health, Nagoya, Aichi, Japan

Received 7 May 2004/ Returned for modification 14 July 2004/ Accepted 13 August 2004

We developed a rapid PCR method utilizing the diversity of the insertion site IS1203 for genotyping Shiga toxin-producing Escherichia coli (STEC) O157 (IS1203 PCR typing). DNA fragments digested by PvuII, which cut IS1203 at one site, were ligated with themselves and detected by PCR with outward-facing primer pairs for IS1203. To minimize nonspecific bands, nested PCR was also performed. Two fingerprinting patterns produced from the upstream or downstream regions of IS1203 were obtained within 1 or 2 days. By combining the two patterns, 79 STEC O157 isolates were classified into 39 types, which were then classified into 36 subtypes by pulsed-field gel electrophoresis (PFGE). The discriminatory power of IS1203 PCR typing (D = 0.974) is similar to that of PFGE (D = 0.981). This method can be used for rapid and simplified genotyping.

* Corresponding author. Mailing address: 7-6 Nagare Tsuji-machi Kita-ku, Nagoya 462-8576, Aichi, Japan. Phone: 81-52-910-5669. Fax: 81-52-913-3641. E-mail: masahiro_4_suzuki{at}pref.aichi.lg.jp.

Journal of Clinical Microbiology, December 2004, p. 5462-5466, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5462-5466.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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