钩端螺旋体DNA限制性内切酶图谱鉴定分类的初步研究

PRELIMINARY RESEARCH OF CLASSIFICATION AND IDENTIFICATION FOR LEPTOSPIRA INTERROGANS BY RESTRICTION ENDONUCLEASE ANALYSIS

用ＥｃｏＲⅠ等六种限制性内切酶消化分析了七日热群和波摩那群各型标准株以及一些野生株钩端螺旋体ＤＮＡ。结果表明，不同血清型菌株具有完全不同的限制性内切酶图谱（ＲＥＰ）；来自不同地域、不同时期、不同宿主的同型野生株ＲＥＰ一致，与它们相应的标准株ＲＥＰ没有差别或仅有微小差别。说明钩端螺旋体ＤＮＡ内切酶图谱具有血清型特征性ＲＥＰ，而且相当稳定。本研究测算出七日热和波摩那各型标准株的ＥｃｏＲⅠＲＥＰ五条高分子带的分子量，并以此作为标准鉴定了部分野生苗株。结果显示，仅凭ＥｃｏＲⅠＲＥＰ两条最大的带就可鉴别两群各型菌株。我们认为ＲＥＰ是一种比较灵敏、快速、简便、准确的钩端螺旋体分类方法，所分型别与传统的血清学型别基本一致。

he genomes of reference strains belonging to Hebdomadis and Pomona serogroups and field isolates belongingto serovars pomona and hebdomadis were analyzed with six restriction endonucleases.The results showed obviousdifferences among reference strains. Field isolates belong to the same serovar from different areas,periods and hosts could nont be differented from each other.And the isolates were in accord with their reference strains or there were some tiny differences between them.We attempted to calculate the first five high molecular-weight fragments with EcoR Ⅰ,which were from the reference strains.Some field isolates were identified with these standards.The results showed that strains from Hebdomadis and Pomona serogroups can all be identified only with the first two high molecular-weight fragments.Therefore,REP is considered tentatitively to be more sensitive, specific,accurate.simple and convenient method for classification and identification of Leptospira.And REP types were in accord with serovars.