TY - JOUR
T1 - Development of a rapid PCR method using the insertion sequence IS1203 for genotyping shiga toxin-producing Escherichia coli O157
AU - Suzuki, Masahiro
AU - Matsumoto, Masakado
AU - Hata, Mami
AU - Takahashi, Masao
AU - Sakae, Kenji
PY - 2004/12
Y1 - 2004/12
N2 - 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.
AB - 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.
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U2 - 10.1128/JCM.42.12.5462-5466.2004
DO - 10.1128/JCM.42.12.5462-5466.2004
M3 - Article
C2 - 15583266
AN - SCOPUS:10844226351
SN - 0095-1137
VL - 42
SP - 5462
EP - 5466
JO - Journal of clinical microbiology
JF - Journal of clinical microbiology
IS - 12
ER -