Rapid categorization of pathogenic Escherichia coli by multiplex PCR
Rapid categorization of pathogenic Escherichia coli by multiplex PCR
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DOI：
10.1111/j.1348-0421.2005.tb03752.x
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发表时间：
2005-01-01
影响因子：
2.6
通讯作者：
Nagai, Y
中科院分区：
文献类型：
--
作者：
Kimata, K;Shima, T;Nagai, Y
建立了一步法多重聚合酶链反应（PCR）检测大肠杆菌12个毒力基因的方法。为了区分大肠杆菌的五种类型，本文对大肠杆菌的五种类型进行了分类。大肠杆菌中筛选出肠出血性大肠杆菌的靶基因six 1、stx 2和eaeA。大肠杆菌（EHEC）; coli（EPEC）; invE为肠侵袭性E.大肠杆菌（EIEC）;大肠杆菌（ETEC）; coli（EAggEC）; astA分布于大肠埃希菌的各个种类。杆菌在我们的多重PCR系统中，所有12个靶基因（six 1、stx 2、eaeA、invE、elt、estp、astA、esth、bfpA、aggR、EAF和CVD 432）在一个试管中的单一PCR反应中扩增并通过电泳检测。应用多重PCR技术对208株临床分离的大肠埃希菌进行了检测。大肠杆菌中的毒力质粒的存在，我们的实验室成功地进行了分类和容易分析。
A one-shot multiplex polymerase chain reaction (PCR) was developed for detecting 12 virulence genes of diarrheagenic Escherichia coli. In order to differentiate between the five categories of diarrheagenic E. coli, we selected the target genes: six1, stx2, and eaeA for enterohemorrhagic E. coli (EHEC); eaeA, bfpA, and EAF for enteropathogenic E. coli (EPEC); invE for enteroinvasive E. coli (EIEC); eft, estp, and esth for enterotoxigenic E. coli (ETEC); CVD432 and aggR for enteroaggregative E. coli (EAggEC); and astA distributed over the categories of diarrheagenic E. coli. In our multiplex PCR system, all 12 targeted genes (six1, stx2, eaeA, invE, elt, estp, astA, esth, bfpA, aggR, EAF, and CVD432) were amplified in a single PCR reaction in one tube and detected by electrophoresis. Using our multiplex PCR, the 208 clinically isolated strains of diarrheagenic E. coli in our laboratory were successfully categorized and easily analyzed for the presence of virulence plasmids.