斯氏狸殖吸虫成虫可溶性抗原蛋白质分析  

ANALYSIS OF SOLUBE ANTIGENS OF PAGUMOGONIMUS SKRJABINI

作　　者：陈文碧[1] 张锡林[2] 王光西[1] 许颖[2] 张跃辉[1] 佘俊萍[1] 杨兴友[1] 

机构地区：[1]泸州医学院病原生物学教研室,四川泸州646000 [2]第三军医大学基础部病原生物学教研室,重庆400038

出　　处：《中国寄生虫病防治杂志》2005年第6期422-425,共4页Chinese Journal of Parasitic Disease Control

基　　金：四川省教育厅科研重点资助项目(No.2005A067)

摘　　要：目的分析和检测斯氏狸殖吸虫成虫可溶性抗原的特异抗原组分,为建立特异、敏感的免疫学诊断方法、研制抗斯氏狸殖吸虫药物和疫苗提供理论基础. 方法用聚丙烯酰胺凝胶电泳(SDS-PAGE)、双向电泳(Two-dimensional gel electrophoresis,2-DE) 和免疫印迹(Western blot)等方法,对斯氏狸殖吸虫成虫可溶性抗原进行分析. 结果 SDS-PAGE分离出22条蛋白带, 其中主带8条,分子质量单位为13～64 ku,未见65 ku以上条带.Western blot显示20条显色带, 主带6条,分子质量单位分别为 13、18、22、28、35 和 64 ku.2-DE电泳分离出斯氏狸殖吸虫成虫可溶性抗原48个多肽斑点, 分子质量单位为14～70 ku,绝大部分分布于pI 3.10～7.14,少数分布于pI 8.78～9.85,在 pI 7.14～8.78之间几乎未见多肽斑点.Western blot分析出其中的10个主要多肽斑点能被斯氏狸殖吸虫病患者血清识别,大部分为集中在酸性区域的小分子多肽. 结论斯氏狸殖吸虫成虫可溶性抗原经SDS-PAGE电泳分离出的22条蛋白带中,有6条蛋白含量较高,能被斯氏狸殖吸虫病患者血清识别.2-DE电泳分离出的48个多肽斑点中,有10个含量较高,能被斯氏狸殖吸虫病患者血清识别,大部分为偏酸性的小分子多肽.Objective To analyze the soluble antigens of Pagumogonmus skrjabini adult worm and demonstrate the specific antigens, Methods 2 dogs were infeeted with metaeercarias of P, skrjabini eolleeted from erab in Xingwen County of Siehuan. After 3 months infection, the adult worms were isolated from infected dog. The soluble antigens of adult worm were analyzed by SDS PAGE, two-dimensional gel electrophoresis（2-DE） and western blot. Results Using 10% resolution gel and 5% stacking gel, a total of 22 distinct protein bands of the soluble antigens of adult worm were demonstrated by SDS-PAGE after Coomassie Blue R-250 stain. Eight of them were main bands, with MWs 13--64 ku. By western blot, sera of humans infected by P. skrjabini recognized out 20 protein bands with 6 main bands in them. Two-dimensional gel eleetrophoresis revealed up to 48 individual polypeptidespots in the soluble antigens of adult worm. The pI and MWs of these spots were 3.10--7.14 and 14--70 ku, respectively. By western blot, sera of humans infected by P. skrjabini recognized 10 main polypeptide spots among them. Conclusion 6 protein bands out of 2Z soluble antigen bands of the parasite showed high immunogenieity. 2-DE revealed up to 48 individual polypeptide spots in the soluble antigens of adult worm. By western blot, sera of humans infected by P. skrjabini recognized 10 main polypeptide spots among them.

关 键 词：斯氏狸殖吸虫 抗原 SDSPAGE免疫印迹 双向电泳 

分 类 号：R383.29[医药卫生—医学寄生虫学]