双抗夹心—ELISA诊断牛隐孢子虫病  

DIAGNOSIS OF CRYPTOSPORIDIOSIS IN BOVINE BY A SANDWICH-ELISA

作　　者：尹继刚[1] 张西臣[1] 陈建宝 李建华[1] 杨举[1] 田宗成 李德昌[1] 

机构地区：[1]解放军农牧大学动物医学系寄生虫病教研室

出　　处：《中国兽医寄生虫病》1999年第2期1-3,共3页Chinese Journal of Veterinary Parasitology

基　　金：军队医药卫生青年基金

摘　　要：为了建立双抗体夹心—ＥＬＩＳＡ检测牛粪便中隐孢子虫卵囊抗原的方法。采用抗小球隐孢子虫（Ｃ．ｐａｒｖｕｍ）卵囊壁单克隆抗体，经对６０头份牛粪便样本分别进行抗酸染色和双抗夹心—ＥＬＩＳＡ检测，结果抗酸染色法检出１２头份有隐孢子虫卵囊，而ＥＬＩＳＡ除对抗酸染色阳性的１２份粪样判为阳性外，还对抗酸染色阴性的４份粪样判为阳性，且不与牛球虫、牛结肠小袋纤毛虫发生类属反应。此外，本试验在稀释液中加入ＥＤＴＡ，并增加了反应温度，使得试验在抗体包被板并封闭后３０ｍｉｎ结束整个检测过程。结果表明，双抗体夹心—ＥＬＩＳＡ是敏感性高、特异性强的诊断牛隐孢子虫病的方法。A sandwich ELISA was established to detect Cryptosporidium oocyst antigen in bovine feces. A monoclonal antibody against C.parvum oocyst wall were used in the experiment.A total of 60 feces samples were detected by acid-fast stain and sandwich-ELISA respectively.The results indicated that 12 feces samples were positive by both acid-fast stain and sandwich-ELISA.However 4 negative feces samples determined by acid-fast stain were determined as positive by sandwich-ELISA.Cross-reaction indicated that feces with Eimeria spp or Balantidium spp were negative by sandwich-ELISA.With the addition of EDTA in diluting solution and increation of reactive temprature,the feces samples were detected within 30 min by sandwich ELISA after coatting and blocking.These results indicated that the sandwich-ELISA was more sensitive and specific for diagnosis of cryptosporidiosis in bovine.

关 键 词：隐孢子虫病 诊断 双抗体夹心-ELISA 

分 类 号：S858.235.9[农业科学—临床兽医学]