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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 ,这在钩体病分子流行病学研究中具有重要意义
Objective To establish the 15 strains of Leptospira interrogans standard strains of Leptospira strain 15 in China by means of pulsed-field gel electrophoresis and to identify the pathogenic leptospira. Methods The whole chromosomal DNA of Leptospira bacterium was directly purified by low melting point agarose gel block and digested with restriction endonuclease NotⅠ and then separated by pulsed-field gel electrophoresis. Results The DNA fragments from 50 to 100 kb in size were well separated, and 15 domestic 15 standard strains of domestic strains were characterized by pulsed-field gel electrophoresis. Conclusion Pulsed-field gel electrophoresis can be used to classify and identify Leptospira bacteria, which is of great significance in molecular epidemiology of leptospirosis