生物素标记ST基因探针检测产肠毒素性大肠杆菌的研究

Detection of Enterotoxigenic Escherichia coli with Biotinylated ST Gene Probe

采用二步法缺口翻译反应,用Bio-11-dUTP标记编码大肠杆菌耐热肠毒素的DNA片段作为杂交探针,通过菌落杂交试验,在严格去蛋白和RNA的条件下,表明生物素标记的DNA探针对产肠毒素性大肠杆菌的检测具有高度特异性.用链亲和素和生物素-碱性磷酸酶系统,通过点杂交试验可以检测出1.25pg的靶序列DNA.

DNA encoding a heat-stable enterotoxin of Escherichia coli was label ledby two-step nicktranslation with Bio-11 -dUTP residues. The biotinylated DNA probe was highly specific for the detection of Enterotoxigenic Escherichia coli when reacted with protein-and RNA-free DNA in a colony hybridization assay. As little as 1.25pg of target DNA can be visualized using dot blot hybridization procedure in conjunction with streptavidin and biotinylated alkaline phosphatase. The DNA hybridization assay with biotinylated DNA probe is efficient for detection of Enterotoxigenic Escherichia coli.