鹅副黏病毒NP蛋白间接ELISA检测方法的建立及免疫鹅抗体水平的检测  

Development of an indirect-ELISA assay based on NP protein of goose paramyxovirus and detection of antibody levels in immunized geese

作　　者：周丽[1] 藏玉婷[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2007年第8期695-699,共5页Chinese Veterinary Science

基　　金：黑龙江省"十五"科技攻关项目(GB01B503-02)

摘　　要：以纯化的鹅副黏病毒NP蛋白为包被抗原,建立了检测鹅副黏病毒NP蛋白抗体的间接ELISA方法,并确定了间接ELISA的最适反应条件:抗原包被浓度为1.0μg/mL,血清稀释度为1∶200,兔抗鹅IgG辣根过氧化物酶标记抗体稀释度为1∶2 000,抗原和血清、血清和二抗均于37℃反应1 h,底物于37℃显色15 min。此间接ELISA方法的特异性强、重复性好。应用该方法对试验鹅血清进行检测,以HI试验为参照。经统计学处理后,比较了两方法测得的抗体效价,分别建立了各组鹅群的回归方程。显著性检验证明,这两种方法检测的抗体效价呈显著相关关系。Using the purified recombinant nucleocapsid protein（NP）of goose paramyxovirus（GPMV） as coating antigen,an indirect-ELISA assay was developed for the detection of antibody against GPMV. In the indirect-ELISA, the optimal concentration of the recombinant NP for coating was 1μg/mL, the optimal dilution of the rabbit anti-goose IgG labeled with HRP was 1 ： 2 000,the optimal dilution of serum sample was 1：200,the serum sample and the HRP-labeled rabbit anti-goose IgG should be incubated at 37 ℃ for 1 h,and the substrate should be incubated at 37 ℃ for 15 rain. The indirect-ELISA assay was confirmed to have good reproducibility and specificity. Goose serum samples were detected by the indirect-ELISA and HI test,and the titers of the antibody were compared between the two methods. The regression equation was developed between titers of ELISA and HI test in each experiment group of geese. The significance testing demonstrated that the titers detected by the indirect-ELISA were correlated significantly with those detected by the HI test.

关 键 词：鹅副黏病毒 NP蛋白 间接ELISA 消长规律 

分 类 号：S852.659[农业科学—基础兽医学]