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首页> 外文期刊>Journal of Clinical Microbiology >Multiplex PCR Genotyping Assay That Distinguishes between Isolates of Clostridium perfringens Type A Carrying a Chromosomal Enterotoxin Gene (cpe) Locus, a Plasmid cpe Locus with an IS1470-Like Sequence, or a Plasmid cpe Locus with an IS1151 Sequence
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Multiplex PCR Genotyping Assay That Distinguishes between Isolates of Clostridium perfringens Type A Carrying a Chromosomal Enterotoxin Gene (cpe) Locus, a Plasmid cpe Locus with an IS1470-Like Sequence, or a Plasmid cpe Locus with an IS1151 Sequence

机译：多重PCR基因分型分析，可区分携带细菌肠毒素基因（cpe）基因座，具有IS1470序列的质粒cpe基因座或具有IS1151序列的质粒cpe基因座的产气荚膜梭状芽孢杆菌分离株

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Clostridium perfringens type A isolates carrying the enterotoxin (cpe) gene are important causes of both food poisoning and non-food-borne diarrheas in humans. In North America and Europe, food poisoning isolates were previously shown to carry a chromosomal cpe gene, while non-food-borne gastrointestinal (GI) disease isolates from those two geographic locations were found to have a plasmid cpe gene. In this report, we describe the development of an economical multiplex PCR cpe genotyping assay that works with culture lysates to distinguish among type A isolates carrying a chromosomal cpe gene, a plasmid cpe gene with a downstream IS1470-like sequence, or a plasmid cpe gene with a downstream IS1151 sequence. When this multiplex PCR assay was applied in molecular epidemiologic studies, it was found that (i) all 57 examined type A isolates with a plasmid cpe gene have either IS1470-like or IS1151 sequences downstream of the plasmid cpe gene; (ii) an IS1470-like sequence, rather than an IS1151 sequence, is more commonly present downstream of the plasmid cpe gene (particularly in North American non-food-borne human GI disease isolates); and (iii) as previously shown in the United States and Europe, isolates carrying the chromosomal cpe gene also appear to be the major cause of C. perfringens food poisoning in Japan. The superiority of this new multiplex PCR assay over existing cpe genotyping approaches should facilitate further molecular epidemiologic investigations of C. perfringens enterotoxin-associated GI illnesses and their associated cpe-positive type A isolates.
机译：携带肠毒素（ cpe ）基因的产气荚膜梭状芽孢杆菌（A）分离株是人类食物中毒和非食物性腹泻的重要原因。在北美和欧洲，以前显示食物中毒分离株带有染色体 cpe 基因，而发现来自这两个地理位置的非食物源性胃肠道（GI）疾病分离株具有质粒< em> cpe 基因。在这份报告中，我们描述了一种经济的多重PCR cpe 基因分型测定法的发展，该法与培养物裂解物一起区分带有染色体 cpe 基因的A型分离株，质粒<带有IS 1470 下游序列的em> cpe 基因，或带有IS 1151 下游序列的质粒 cpe 基因。当将此多重PCR分析应用于分子流行病学研究时，发现（i）所有57个带有质粒 cpe 基因的A型分离株都具有IS 1470 样质粒 cpe 基因下游的IS 1151 序列; （ii）类似于IS 1470 的序列，而不是IS 1151 序列，更常见于质粒 cpe 基因的下游（尤其是在北美非食源性人类胃肠道疾病分离物中）; （iii）如先前在美国和欧洲所示，带有染色体 cpe 基因的分离株也似乎是 C的主要原因。日本发生perfringens 食物中毒。与现有的 cpe 基因分型方法相比，这种新的多重PCR测定法的优越性应有助于 C的进一步分子流行病学研究。产气荚膜炎与肠毒素有关的胃肠道疾病及其相关的 cpe 阳性A型分离株。

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