不同毒力钩端螺旋体属特异性外膜蛋白抗原的分析

Analysis on the genus-specific antigens from outer membrane protein of Leptospira with different virulence

目的 了解不同毒力钩端螺旋体株有无属特异性外膜蛋白抗原。方法 TR/ patocⅠ钩体属特异性抗原免疫家兔获得抗血清 ,以显微镜凝集试验检测TR/patocⅠ属特异性抗原抗血清对我国 15群 17型问号钩体和 2群 2型双曲钩体的凝集情况。采用Auran介绍的方法制备问号钩体黄疸出血群赖型 5 6 6 0 1株、波摩那群波摩那型 5 6 6 0 8株和双曲钩体三宝垄群Patoc型patocⅠ株的外膜蛋白 ,用SDS -PAGE及Westernblot分析不同毒力钩体外膜蛋白的电泳特征及与TR/ patocⅠ属特异性抗血清的反应性。结果 钩体TR/ patocI属特异性抗血清能与上述各株钩体发生凝集反应 ,其效价为 1∶2 5 6～ 1∶5 12。三种不同毒力钩体外膜蛋白有相似的SDS -PAGE图谱 ,其主要蛋白组分的分子量约为 6 0kDa ,Westernblot结果证实在约31kDa处有一共同的能与TR/patocI属特异性抗血清发生反应的阳性条带。结论 钩体细胞表面具有属特异性抗原 ,分子量31KDa外膜蛋白可能是属特异性抗原之一。

To understand the existence of genus-specific antigens from outer membrane proteins (OMP) of Leptospira with different virulence,rabbit antiserum against leptospiral TR/patoc I antigen was prepared for microscope agglutination test (MAT).MAT was applied to detect the agglutination of 17 strains of Leptospira interrongans of 15 serogroups and 2 strains of Leptospira bifrexa of 2 serogroups with the antiserum.The OMPs of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601,serogroup Pomona serovar pomona strain 56608 and L.biflexa serogroup Semaranga serovar patoc strain patoc Ⅰ were prepared by using the method introduced by Auran et al.SDS-PAGE and western blotting were used for analyzing the features of electrophoresis and immunoreactivity to antiserum against TR/patoc Ⅰ genus-specific antigen,respectively.It was found that all the strains belong to different leptospiral serogroups could react with the antiserum against TR/patoc I genus-specific antigen and the agglutination titers were ranged from 1∶256～1∶512.A similar SDS-PAGE map of the OMPs from the three strains of leptospira with different virulence was present and molecular Weights of the major protein fragments were approximate 60kDa.The result of western blotting showed that a protein fragment with 31kDa from each of the three strains was able to react with antiserum against TR/patoc Ⅰ genus-specific antigen.It condudes that there are genus-specific antigens on the surface of leptospiral bodies.The OMP with molecular weight 31kDa may be one of the genus-specific antigens.