检测ＮＤＶ特异性ＩgG、IgM、ＩgA的间接ＥＬＩＳＡ方法的建立

本试验以新城疫病毒（ＮＤＶ）感染的鸡胚尿囊液经差速离心，庶糖密度梯度离心提纯的ＮＤＶ为包被抗原，以纯化的鸡ＩgG、IgM及ＩgA免疫，制备辣根过氧化物酶标记的兔抗鸡ＩgG、IgM及ＩgA，分别建立了检测ＤＮＶ特异性ＩgG、IgM、ＩgA的间接ＥＬＩＳＡ方法，抗原最适包被量为１．３４μg/孔。酶标兔抗鸡ＩgG、IgM、ＩgA的最佳稀释度分别为１：４００，１：４００，１：１００，血清样品检测ＩgG时，最适工作浓度为１：４００，检测IgM时为１：５０。该方法具有特异性强、灵敏度快，快速简便等优点，适合于对各类新城疫特异性抗体的动态监测和大批检测，同时也为城疫免疫要理研究及流行病学调查提供了可靠手段。

Indirect ELISA for theDetection of NDV Specific Immunoglobulins G,M

Indirect ELISAs for estimation of Newcastle disease(ND)virus specific IgG、IgM、IgA were developed by using purified NDV antigen and rabbit antichicken IgG、IgM、IgA conjugated with horseradish peroxides(HRP),the whole procedure was optimized.Polystyrene chloride microtitration plates were coated with purified NDV antigen at the amount of 1.37μg in 100μl 0.05M Sodium bicarbonate buffer(pH9.6)per well.The working concentration of HRP_labelled rabbit antichicken IgG,IgM，and IgA was 1∶400,1∶400,1∶100,and that of serum samples respectively was 1∶400 and 1∶50 in detection of IgG and IgM.Those ELISA systems are sensitive and specific,are suitable techniques for the assay and surveillance of chicken NDV specific IgG,IgM,IgA,can be used in the investigation of chicken immune mechanism.