亲和层析法纯化华支睾吸虫抗原的研究

目的探索华支睾吸虫特异性抗原的纯化方法。方法从华支睾吸虫病流行区的淡水鱼中分离华支睾吸虫囊蚴,接种动物,收集成虫,制备匀浆液(粗抗原),借助偶联华支睾吸虫病人血清抗体的Sepharose4B亲和层析柱,提取特异性抗原,用浓度梯度聚丙烯酰胺凝胶电泳(CG-PAGE)和免疫印迹方法鉴定纯化抗原的纯度和特异性。结果亲和层析法纯化抗原经CG-PAGE显示5条蛋白带,免疫印迹试验鉴定均有抗原性,强阳性带蛋白分子质量单位为31ku,各区带蛋白与并殖吸虫、血吸虫血清抗体无交叉反应。粗抗原有16条蛋白带。结论经亲和层析法纯化的华支睾吸虫抗原特异性高。亲合层析是获取华支睾吸虫诊断用抗原的较佳方法。

STUDY ON THE PURIFICATION OF CLONORCHIS SINENSIS ANTIGEN BY AFFINITY CHROMATOGRAPHY

Objective To explore the purification methods of the Clonorchis sinensis antigen. Methods Metocecarias were separated from the fish in clonorchiasis endemic area and infected animals. The adult worms were collected from the infection animals and then homogenates were made, in which, specification antigen were isolated by means of affinity chromatography with Sepharose 4B linked serum antibody from patients with clonorchiasis, then identified by CG-PAGE and Western blot. Results The purified antigen has 5 bands and 31 ku protein showed high antigenic, no cross reaction with Paragonimus and Schistosoma antibodies; 16 bands appeared in unpurified antigen. Conclusion Affinity chromatography is good methods for separated diagnostic antigen from the worm homogenates.