斯氏狸殖吸虫童虫、成虫排泄分泌产物的分析研究

Analysis of Excretory-secretory Products of Adult and Larva of Pagumogonimus skrjabini

目的分析斯氏狸殖吸虫童虫、成虫排泄分泌产物(ES)的蛋白条带和抗原性,并对两者进行免疫组织定位。方法十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离斯氏狸殖吸虫童虫ES蛋白、童虫虫体可溶性蛋白、成虫ES蛋白、成虫虫体可溶性蛋白,采用Western-blot方法分别用肺吸虫病人血清、小鼠抗血清识别特异性反应条带,并用酶标免疫组织化学技术对ES抗原进行组织定位。结果经SDS-PAGE分离的斯氏狸殖吸虫的虫体可溶性蛋白和ES蛋白,Western-blot结果显示,感染血清识别的童虫ES蛋白、成虫ES蛋白条带分别在28000、70000Mr和90000Mr处,小鼠抗血清识别的童虫ES蛋白、成虫ES蛋白条带分别在28000、70000Mr和45000、70000Mr处。童虫ES主要定位在后尾蚴的排泄囊,成虫ES主要定位在成虫肠腔上皮。结论 28000、70000Mr的童虫ES蛋白可能为斯氏狸殖吸虫的特异诊断抗原。

Objective To analyze the immunological properties of the excretory-secretory products（ES） produced by Pagumogonimus skrjabini.Methods ES and soluble proteins from the adult and larva of Pagumogonimus skrjabin were analyzed by Western-blot using both patient and mouse immune serum.HRP immunohistochemical technique was used to determine the localization of the excretory-secretory antigens in the adult worm.Results Using the patient immune serum,the molecular weight of larval ES proteins was found to be 28 000 and 70 000 Mr,and the adult ES protein was 90 000 Mr.Using the immune mouse serum,the molecular weight of larval ES proteins was found to be 28 000 and 70 000 Mr,and the adult ES protein was 45 000 and 70 000 Mr.Larval ES was localized mainly in the bladder of the metacercaria,while the adult ES was localized mainly in the intestinal epithelium of the adult worm.Conclusion ES proteins with the molecular weight of 28 000 and 70 000 Mr be used for clinical diagnosis.