家兔黄艾美耳球虫血清抗体ELISA检测法的建立

Establishment of an Enzyme-linked Immunosorbent AssayUsed for Detecting Serum Antibody in Rabbit with Coccid-iosis

应用制备的孢子化卵囊可溶性抗原作为包被抗原建立了兔球虫血清抗体的ＥＬＩＳＡ检测法。其最适条件为：抗原包被浓度，２００μｇ／ｍＬ；包被温度，３７℃（２ｈ）；血清反应时间，１．５ｈ；酶标抗体标准工作浓度，１：１０００；反应时间，１ｈ；待检血清稀释度，１：１００；ＥＬＩＳＡ试验的规定吸收值为０．０６。孢子化卵囊可溶性抗原比非孢子化卵囊可溶性抗原更适于作为ＥＬＩＳＡ包被抗原。

An enzyme-linked immunosorbent assay (ELISA ) for detecting serumantibody in rabbit with coccidiosis was successfully established.The soluble antigen wasprepared from sporulated oocysts of Eimeria flavescens. The optimal parameters for theassay were as follows :200 μg/mL of soluble antigen was coated for 2 h at 37℃; antis-era were incubated for 1.5 h and u:ed at dilution of 1:100; enzyme conjugate was usedat dilution of 1 :1 000,incubated for 1 h. Pre-set positive absorbance value was 0.06and the specificity tests were performed. As compared with soluble antigen from un-sporulated oocysts,the soluble preparation from sporulated oocysts was more suitablefor ELISA as a coating antigen。