title: Development of a rapid PCR method using the insertion sequence IS1203 for genotyping Shiga toxin-producing Escherichia coli O157.
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-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.
authors.affiliation: Department of Microbiology, Aichi Prefectural Institute of Public Health, Nagoya, Aichi, Japan. masahiro_4_suzuki@pref.aichi.lg.jp
authors.forename: Masahiro
authors.initials: M
authors.lastname: Suzuki
authors.forename: Masakado
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Mami
authors.initials: M
authors.lastname: Hata
authors.forename: Masao
authors.initials: M
authors.lastname: Takahashi
authors.forename: Kenji
authors.initials: K
authors.lastname: Sakae
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology*; D004251:DNA Transposable Elements; D016521:Electrophoresis, Gel, Pulsed-Field; D004927:Escherichia coli Infections / Q000175:diagnosis / Q000382:microbiology* / Q000662:veterinary; D019453:Escherichia coli O157 / Q000145:classification* / Q000235:genetics; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D022603:Shiga Toxins / Q000096:biosynthesis; D013997:Time Factors
chemical_list: D004251:DNA Transposable Elements; D022603:Shiga Toxins