PCR在大肠杆菌ST1b基因扩增及其基因检测上的应用

The Application of Polymerase Chain Reaction (PCR) in Am plication and Detection of STlb Gene of ETEC

作者应用聚合酶链反应(PCR)成功地扩增了产肠毒性大肠杆菌ST1b基因,在ST1b产量、活性和纯度上均满足了基因重组的要求。并利用PCR技术建立了一种新型基因探针标记方法(基因扩增标记法),所制备的ST1b基因探针经相关菌菌落杂交试验证实具有良好的特异性和敏感性。通过对180株婴幼儿腹泻大肠杆菌分离株、52株仔猪腹泻大肠杆菌分离株和28株大肠杆菌标准菌株的基因诊断以及96株K99—ST1b基因重组转化菌的基因检测,菌落杂交阳性率分别为11/180、2/52、2/28和5/96。

The STlb gene of ETEC was amplified successfully by PCR technology, and the quantity, activity and purity of the STlb gene obtained all achieved the requirement of the gene recombination. A new method for gene probe labeling (gene Amplifing-labeling) was established by PCR. The probe has better specificity and sensitivity when identified by clony hybridization in situ with relative bacteria. 180 strains of E. coli which isolated from children diarrhea, 52 strains of E. coli from pigletdiarrhea, 28 standard E. coli strains which serum types have been known, and 96 strains of re-combinant clones were detectea using STlb geng probe. The hybridization positive rate was 11/180, 2/52, 2/28 and 5/96. respectively.