结核分枝杆菌H_(37)Rv株菌体抗原及分泌抗原分析

Analysis of cultured and secreted antigens in Mycobacterium tuberculosis H_(37)Rv

目的对比分析人型结核枝杆菌(结核菌)H37Rv株固体培养菌、液体培养菌及分泌蛋白中抗原成分的差异,寻找结核菌的主要免疫抗原。方法应用SDS聚丙烯酰胺凝胶电泳技术、免疫印迹技术及单克隆抗体技术分析人型结核菌H37Rv株固体培养、液体培养及其分泌蛋白中的抗原成分。结果固体培养菌与液体培养菌具有基本一致的多肽抗原成分,其主要成分分子量为79000、64000、54000、50000、28000～38000、23000等,而分泌蛋白差异稍大,其主要成分分子量为66000、63000～65000、55000、42000、32000～34000、24000、16000等。单克隆抗体进一步证明66000、55000、16000为分泌蛋白。多克隆抗体免疫印迹分析表明,菌体主要免疫原有79000、54000、38000～50000、28000蛋白成分,而分泌蛋白主要免疫原为分子量66000、55000～64000、30000～34000、24000、16000成分。结论人型结核菌H37Rv株菌体蛋白和分泌蛋白具有不同的主要免疫原,将有助于认识HRv株的总体蛋白组成及寻找与结核病相关的特异性抗原成分。

Objective To analyze the antigens of solid cultured germ, liquid cultured germ and secreted composition, looking for the main immunity antigens of Mycobacterium tuberculosis H37 Rv. Methods Showing the difference of the antigens of solid cultured germ, liquid cultured germ and secreted composition by SDS-PAGE, Western blotting, monoclonal and polyclonal antibody techniques. Results Antigens of the solid cultured germ were nearly accordance with the liquid cultured germ and they were difference with secreted proteins. The main immunity antigens of the cultured germ were 79 000, 54 000 ,38 000～50 000,28 000 and the secreted proteins are 66 000, 55 000～ 64 000, 30 000 ～ 34 000, 24 000, 16 000. The antigens of 66 000, 55 000 and 16 000 were proved that they were secreted proteins by monoclonal antibodies. Conclusions This study demonstrated that the main immunity antigens of the cultured germ in Mycobacterium tuberculosis H37 Rv were difference with the secreted one . This is beneficial to the cognition of H37 Rv and looking for special antigens for diagnosis of tuberculosis .