致病性钩端螺旋体脉冲场凝胶电泳图谱分型研究

Identification of Leptospira interrogans by pulsed-field gel electrophoresis

目的 建立我国 15群 15型钩端螺旋体标准菌株染色体DNA脉冲场凝胶电泳图谱 ,进行致病性钩体的分子遗传学分类鉴定。方法 采用低熔点琼脂糖凝胶块直接纯化钩体菌完整的染色体DNA ,用限制性内切酶NotⅠ消化后 ,进行脉冲场凝胶电泳分离。结果 从 5 0～ 10 0 0kb范围的DNA片段得到很好的分离效果 ,我国常见 15群 15型国内标准菌株各具特异性脉冲场凝胶电泳图谱。结论 脉冲场凝胶电泳图谱可对钩体菌进行遗传学分类鉴定 。

Objective To establish a genetic typing maps for Leptospira interrogans domestic reference strains by pulsed field gel electrophoresis (PFGE). Methods Low melting point agarose plug was used to purify the intact chromosome DNA from Leptospira intenogans . The chromosome DNAs were digested with restriction endonuclease NotⅠand the fragments were separated by PFGE. Results The DNA fragments from 50～1?000kb were separated clearly, each of the 15 domestic reference strains showed a specific PFGE pattern. Conclusion PFGE patterns, which could be used to classify and identify Leptospira interrogans , could be used in molecular epidemiological research of leptospirosis.