Luo Yi-hui, Yan Jie, Mao Ya-fei, Li Shu-ping
Department of Medical Microbiology and Parasitology, College of Medical Science, Zhejiang University, Hangzhou 310006, China.
J Zhejiang Univ Sci. 2004 Apr;5(4):462-6. doi: 10.1631/jzus.2004.0462.
To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) of leptospira with different virulence.
Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains of Leptospira interrongans belonging to 15 serogroups and 2 strains of Leptospira biflexa belonging to 2 serogroups. The outer envelopes (OEs) of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain lai (56601) with strong virulence and serogroup Pomona serovar pomona strain Luo (56608) with low virulence, and L.biflexa serogroup Semaranga serovar patoc strain Patoc I without virulence were prepared by using the method reported in Auran et al.(1972). OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively.
All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256-1:512. A similar SDS-PAGE pattern of the OMPs from the three strains of leptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 KDa. A positive protein fragment with approximately 32 KDa confirmed by Western blot, was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains of leptospira.
There are genus-specific antigens on the surface of L.interrogans and L.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens of leptospira.
确定不同毒力钩端螺旋体外膜蛋白（OMPs）中属特异性抗原的存在情况。
应用显微镜凝集试验（MAT）检测商业兔抗钩端螺旋体属特异性TR/Patoc I抗原血清与15个血清群的17株问号钩端螺旋体及2个血清群的2株双曲钩端螺旋体之间的凝集反应。采用Auran等人（1972年）报道的方法制备强毒的出血性黄疸群赖型钩端螺旋体赖株（56601）、低毒的波摩那群波摩那型钩端螺旋体罗株（56608）以及无毒的双曲钩端螺旋体塞马朗群帕托克型钩端螺旋体帕托克I株的外膜（OEs）。通过脱氧胆酸钠处理从OEs中获得OMPs。分别使用SDS-PAGE和蛋白质印迹法分析OMPs的电泳图谱特征及其与抗TR/Patoc I抗原血清的免疫反应性。
所有受试的不同钩端螺旋体血清群菌株均与抗钩端螺旋体属特异性TR/Patoc I抗原血清发生凝集，凝集效价范围为1:256 - 1:512。显示出三株不同毒力钩端螺旋体的OMPs具有相似的SDS-PAGE图谱，且发现OMPs中一个主要蛋白片段的分子量约为60 kDa。经蛋白质印迹法确认的约32 kDa的阳性蛋白片段能够与抗钩端螺旋体属特异性TR/Patoc I抗原血清发生反应，并且在三株钩端螺旋体的每个OMPs中均能检测到。
问号钩端螺旋体和双曲钩端螺旋体表面存在属特异性抗原。分子量为32 kDa的OMP可能是钩端螺旋体的属特异性蛋白抗原之一。
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