问号钩体血清型参考株16S　rRNA基因的PCR－SSCP分析

16S rRNA Gene PCR-SSCP Analysis of the Reference Strains from 15 Serovars(14 Serogroups)of Pathogenic Leptospires in China

利用１６ＳｒＲＮＡ基因引物扩增了我国致病性钩体１４群１５型参考株及赖型０１７株ＤＮＡ，通过非变性聚丙烯酰胺凝胶电泳，对各参考株的ＰＣＲ产物单链构象多态性进行了对比。结果显示在不同浓度凝胶及电泳条件下，１６株产物均有明显的２条单链，其中爪哇型、拜伦型、塔拉索夫型和曼耗Ⅱ型的国内参考株有相同的带型，而另１１型１２株的带型一致。此结果与Ｙａｓｕｄａ等（１９８７）和Ｒａ－ｍａｄａｓｓ等（１９９２）遗传学分种基本相符。

The DNAs of reference strains from 15serovars(14 serogroups) of pathogenic leptospires inChina were amplified with 16S rRNA gene primers,and then single-strand conformation polymorphism(SSCP)of the products were analyzed in 12.5%non-denaturing mini polyacrylamide gel(containing 5%glycerol)combining with silver-stainIng. All productsshowed two bands on electrophoresis at different pa-rameters of voltage or current and concentration ofgel.It proved that serovar lai,serovar canicola,serovar pyrogenes,serovar autumnalis, serovar aus-tralis,serovar pomona,serovar linhai,serovar hebdo-madis, serovar haemolytica,serovar wolffi and serovar paidjan have the identical pattern(Leptospirainterrogans),while serovar javanica,serovar ballum,serovar tarassovi and serovar manhao Ⅰ belong to an-other pattern(L.borgpetersenii).The result was con-sistent with the classification of genetic species by Ya-suda et al (1987)and Ramadass et al (1992).