ELISA双抗夹心法检测IBDV抗原

采用IBD细胞毒油佐剂灭活苗多次免疫易感鸡，制取高免血清效价1：64，提纯IgG抗体并用HRP标记同一IgG抗体，建立一种检测IBDV的敏感性方法──ELISA双抗夹心法。实验用盐析法和离子交换层析法纯化IgG。酶标抗体E／IgGmol比1：43。方阵实验确定最佳反应条件；包被抗体50ug／ml，E－Ab2稀释度为1：120，Ag灵敏度为1：3200；取代反应，抗原阻断反应，无关病毒对照试验均为阴性，并与AGP法对照实验．灵敏度高200倍。以上结果表明：本实验敏感性高，特异性强，简单快速判定客观，对临床诊断IBD流行病有很好的使用价值和推广价值。

Using ELISA Douber Antibody Sandwich to Detect IBD Antigens

Vaccine killed by IBD cell adjuvant of virus and oil was used to immune sensitive chickens for several times,and serum with high immunity(the potency is 1: 64)was isolated.IgG was purified and labelled with HRP. The method of ELISA douber antibooly sandwich-a sensitive method to detect IBDV was created. IgG was purified by method of salting out and ion exchange layering out. (The antibodies labelled with enzyme E/IgG moltt 1:43). The perfect reaction conditions determined by Fang and zhen are(envelope antibodies 50 ug/ml),and that the degree of dilution of EAb2 is 1:120,and that the sensitivity of Ag is 1 : 3200. The results of substitution reaction,antigen inhibition reaction and irrelevant control experiment were negative. The sensitivity of the experiment,compared with AGP method,increased by 200 times. The results above demonstrat that the sensitivity and specificity of the experiment are very high,and that the method is simple,fast and objective. Thervfore,it has the values of usage and populazation in detecting IBDG epidemic disease.