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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-dimension-al 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 and detect the specific antigen components of adult soluble antigen of Salexvonchus agglutina, and to provide a theoretical basis for the establishment of specific and sensitive immunological diagnostic methods and the development of anti-Clostridium antigens drugs and vaccines. Methods Soluble antigens of adult Paramisgongi’s moths were analyzed by SDS-PAGE, 2-DE and Western blot . RESULTS: Twenty-two protein bands were separated by SDS-PAGE, of which 8 were major bands with molecular mass units of 13-64 ku. No bands above 65 ku were observed. Western blot showed 20 bands with 6 main bands with molecular mass units of 13, 18, 22, 28, 35 and 64 ku, respectively. 2-DE electrophoresis was isolated from the B. rechsensivi adult worm soluble antigen 48 peptide spots, the molecular mass unit of 14 ~ 70 ku, the vast majority of distribution in the pI 3.10 ~ 7.14, a few distributed in the pI 8.78 ~ 9.85, in the pI 7.14 ~ Few spots of polypeptide were seen between 8.78. Western blot analysis revealed that 10 of the major peptide spots were recognized by the serums of Serratia chinensis, most of which were small molecule peptides concentrated in the acidic region. Conclusion Among the 22 protein bands isolated from SDS-PAGE electrophoresis, the soluble protein of adult Sauropronella minchois has 6 proteins which can be recognized by the serums of the serogroups of Trichoplusia. Among the 48 polypeptide spots isolated by 2-DE, 10 were higher in serum, which could be recognized by the serums of Seriphidium serotina, mostly acidic peptides.