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, Kenji Sakae

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

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-fleld 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.

Original languageEnglish
Pages (from-to)5462-5466
Number of pages5
JournalJournal of clinical microbiology
Volume42
Issue number12
DOIs
Publication statusPublished - 12-2004
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

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