用聚丙烯酰胺凝胶电泳和双向电泳分析阴道毛滴虫滋养体抗原

Antigen Analysis of Trichomonas vaginalis Trophozoite by SDS-PAGE and Two-Dimensional Gel Electrophoresis

目的 研究阴道毛滴虫滋养体可溶性抗原。 方法 用聚丙烯酰胺凝胶电泳 (SDS- PAGE)、酶联免疫电转移印迹 (EITB)和双向电泳等方法 ,对阴道毛滴虫滋养体全虫可溶性抗原进行分析。 结果 SDS- PAGE分离出 2 6条蛋白带 ,其中主带 9条 ,分子量 8条为 15～ 6 2 k Da、1条为 97k Da。EITB显示 2 4条带 ,仅在 75 k Da和2 2 k Da处缺如 ,其中主带 8条。双向电泳分离出 43个多肽斑点 ,多分布于 PI3.6 5～ 5 .84,分子量为 2 7～ >10 0k Da,其中有 9个主要多肽斑点。 结论 阴道毛滴虫滋养体可溶性抗原 2 6条蛋白带中 ,有 8条蛋白含量较高、免疫学反应较强。在 43个多肽斑点中有 9个多肽含量明显较高。

Objective To analyze soluble antigens of Trichomonas vaginalis. \ Methods\ Soluble antigens of the parasite from a patient suffering from trichomonad vaginitis were analyzed by SDS\|PAGE, immunoblotting and two\|dimensional gel electrophoresis. \ Results\ A total of 26 distinct protein bands were demonstrated by using 10% resolution gel. Nine of them were main bands, eight with MWs 15-62 kDa, one with MW 97 kDa. By immunoblotting the specific anti\|T. vaginalis antibodies raised in rabbit recognized 24 protein bands with 8 main bands in them. Two\|dimensional gel electrophoresis revealed up to 43 individual trichomonad polypeptide spots, among which, 9 were main ones. The pI and MWs of these spots were 3\^65-5\^84 and 27->100 kDa respectively. \ \{Conclusion\ Eight\} protein bands out of 26 soluble antigen bands of the parasite showed high immunogenicity. There were 9 main polypeptide spots in 43 polypeptide spots of the parasite.