关键词: 斑点核酸杂交, 钩端螺旋体, 蛙, 流行病学

Abstract:

　　【Abstract】　Objective　To detect leptospira in frogs by Dot blotting and to establish a promising technique used for epidemiological investigation and surveillance of leptospirosis. Methods　A pair of flaB primer was synthesized according to the sequences of flaB from Lai strain of Leptospira icterohaemorrhagiae, and was used to amplify the flaB gene of leptospira in frog specimens collected from leptospirosis epidemic area, Anhui province.  PCR products were then moved to NC membrane and hybridized with flaB probe labeled with digoxiugenin. Results　The results demonstrated that only 5 fg of purified DNA of leptospira could give a positive signal by Dot blotting. There were 19 positive samples detected by Dot blotting among 70 samples of frog kidneys, and the positive rate was 27.14%. Eight samples were positive by cultivating, and the positive rate was 11.43%. A total of 14 samples detected by PCR were positive, and it was 20.00%. Conclusion　Dot blotting by flaB probe labeled with DIG is a high sensitive, specific and rapid technique for epidemiological investigation of frog leptospirosis.

Key words: Dot blotting, Leptospira, Frog, Epidemiology