光敏生物素和^(32)P标记DNA打点杂交检测钩端螺旋体

Detection of Leptospira by Dot Blot Hybridization with Photobiotin- and ^(32)P-labelled DNA

应用光敏生物素和^(32)P标记黄疸出血群赖型017株钩端螺旋体DNA(简称017DNA)作为探针,打点杂交检测问号状钩体。结果表明:两种探针均能检测不同群、型的问号状钩体DNA,最小检测量分别为5pg和1pg;与双曲钩体patoc型PatocⅠ株、细螺旋体属illini型3055株及其他致病菌和正常人血清未发现明显杂交信号。提示光敏生物素制备的017DNA探针可以用于检测钩体,也可应用于钩体病的早期诊断和流行病学调查。

Photobiotin-and ^(32)P-labelledDNA probes of L. interrogans sv. lai strain017 were produced and the DNA and lepto-spires were dotted on the NC filter andhybridized. The results showed that photobiotin and^(32)P-labelled DNA probes could detect DNAsof homology. The smallest amount of DNAthat could be detected with the probes were5pg and 1pg. and the smallest numbers ofpathogenic leptospires were 5×10~? and 10~3,respectively. Nonpathogenic leptospires. L.biflexa sv. pa?oc strain Patoc 1, L. illinistrain 3055, Escherichia coli. Bacillus aero-genes capsulatus, and Salmonella anatis,could not be detected by the probes. Thestudy demonstrates that leptospires DNAprobe could be produced by labelling the DNAwith photobiotin. It has higher sensitivityand specificity and can be used for detec-tion of leptospires in the field and clinic.