文章摘要
蔡庆,王蔚,陈友纯,林万明.三对引物同时PCR分型检测产毒素大肠杆菌的方法及在分子流行病学研究中的应用[J].中华流行病学杂志,1997,18(4):211-213
三对引物同时PCR分型检测产毒素大肠杆菌的方法及在分子流行病学研究中的应用
Study on the Molecular Epidemiology of Diarrhea Caused by Enterotoxigenic Escherichia coli, Using Polymerase Chain Reaction with Multiple Primer Pairs
收稿日期:1997-02-25  出版日期:2021-06-09
DOI:
中文关键词: 产毒素大肠杆菌;腹泻;聚合酶链反应
英文关键词: Enterotoxigenic Escherichia coli(ETEC);Diarrhea;Polymerase chain reaction
基金项目:
作者单位
蔡庆 空军总医院临床分子生物学研究中心 北京 100036 
王蔚 空军总医院临床分子生物学研究中心 北京 100036 
陈友纯 空军总医院临床分子生物学研究中心 北京 100036 
林万明 空军总医院临床分子生物学研究中心 北京 100036 
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中文摘要:
      在产毒素大肠杆菌(ETEC)肠毒素基因内设计合成三对引物,建立了三对引物同时PCR检测ETEC的方法,一次PCR即可扩增出627bp (LTh)、240bp (STIa)和169bp (STIb)三种肠毒素基因片段,可同时分型检测出LTh、STIa、STIb、LTh-STIa、LTh-STIb五种基因型的ETEC,与非ETEC对照菌无交叉反应,最小检出量为10cfu,显示了很高的特异性和敏感性。将建立的方法用于山东省六县市623例大肠杆菌致泻标本的检测,阳性率为40.3%,并能鉴别ETEC的基因型,为ETEC腹泻的分子流行病学研究提供了有效的检测手段。
英文摘要:
      A Polymerase Chain Reaction (PCR) method has been developed for detecting Enterotoxigenic Escherichia coli (ETEC). Three different sets of oligonucleotide primer were simultaneously used to amplify the enterotoxin genes of heat-labile (LTh) and heat-stable (STIa and STIb) enterotoxins of ETEC. These primers amplified the 627, 240 or 169 base pair DNA fragments from LTh, STIa and STIb genes of the reference ETEC strains, respectively.Five types of ETEC strains corresponding to the LTh, STIa,STIb,LTh-STIa,or LTh-STIb genotypes were distinguished by a single procedure of PCR, using the mixture of the three sets of primers.There was no cross-reaction with the non-ETEC strains. The lowest detection level was 10 cfu. A total number of 623 stool specimens of diarrheal patients from Shangdong Province induced by E.coli were examined by PCR and the positive rate of ETEC was 40.3%. The results indicated that PCR is a rapid,sensitive and specific method for detecting ETEC.
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