纤维素滤膜过滤法纯化弓形虫速殖子并制备可溶性抗原  

Purification of Toxoplasma gondii Tachyzoites with Cellulose Membrane Filtration and Preparation of Soluble Antigen

作　　者：方正明[1] 王升[2] 苏斌涛[1] 宫念樵[2] 刘文琪[1] 明长生[2] 

机构地区：[1]华中科技大学同济医学院寄生虫学教研室,武汉430030 [2]华中科技大学同济医学院附属同济医院器官移植研究所,武汉430030

出　　处：《中国寄生虫学与寄生虫病杂志》2010年第5期329-331,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No30772052)~~

摘　　要：取弓形虫RH株感染小鼠腹腔灌洗液20ml,加入无菌生理盐水至250ml,在5μm滤膜孔径的容器过滤器上过滤。收集的滤液经1512×g离心15min,沉淀即为纯净虫体。超声粉碎虫体,11200×g离心30min,收集上清即为弓形虫可溶性抗原。采集感染弓形虫SD大鼠慢性期血清和健康SD大鼠血清,用上述抗原作间接酶联免疫吸附试验(indirectELISA),检测抗原特异性和抗原效价。结果显示,通过改进过滤器、滤膜孔径和过滤方法纯化弓形虫速殖子,平均白细胞清除率为99.9%,红细胞清除率为80.3%,虫体回收率为71.0%。平均每只小鼠提取1.38mg弓形虫可溶性抗原。间接ELISA试验结果表明,抗原效价为5μg/ml。20ml peritoneal lavage fluid of mice infected with Toxoplasma gondii RH strain was diluted to 250ml with sterilized physiological saline, and filtered through cellulose membrane filters （pore size： 5 μm）. The filtrate was centrifuged at 1 512×g for 15 min, and the sediment was pure T. gondii tachyzoites which were then sonicated. The soluble antigen was prepared by centrifugation at 11 200×g for 30 min. Sera of T. gondii infected SD rat and normal SD rats were collected for immunodetection of soluble antigen. The specificity and valence of soluble antigen were detected with indirect ELISA. The mean removal rates of mouse leukocytes and erythrocytes were 99.9% and 80.3%, respectively, and recovery rate of tachyzoites was 71%. The soluble antigen was extracted from purified T. gondii （1.38 mg per mouse）. Indirect ELISA showed that the lowest effective antigen concentration was 5 μg/ml.

关 键 词：弓形虫 纯化 抗原 纤维素滤膜 

分 类 号：R382.5[医药卫生—医学寄生虫学]