猪瘟高免卵黄抗体ELISA检测方法的建立  

Development of indirect ELISA for detecting IgY against SF

作　　者：肖静静[1] 李硕绵[1] 李妹玲 陈立功[1] 王迎春[1] 董世山[1] 

机构地区：[1]河北农业大学动物医学院/河北省兽用生物制品工程技术研究中心,河北保定071001

出　　处：《河北农业大学学报》2013年第5期90-93,112,共5页Journal of Hebei Agricultural University

摘　　要：本试验采用猪瘟活疫苗病毒包被酶标板，提取的卵黄抗体作为一抗，建立针对猪瘟病毒卵黄抗体的间接ELISA检测方法。用方阵滴定法确定各反应液的工作浓度及作用时间，并对检测方法的特异性、敏感性及重复性进行了评估。结果显示，抗原最佳包被浓度为1：1000，酶标二抗工作浓度为1：6000，临界值为0．131～0．145。建立的ELISA方法较IHA敏感性高，用IHA检测效价为2^7的抗体，稀释2^14倍后，用ELISA方法检测仍为阳性，且其特异性及重复性较好。This experiment established an indirect ELISA method for detecting immunoglobulin of yolk（IgY） against swine fever virus （SFV）. The ELISA plates were coated with swine fever vaccine virus, the extracted IgY was used as primary antibody. Optimal concentration of the reaction liquid and action time were determined by phalanx titration method. The specificity, sensibility of antigen-antibody combination and the test repeatability were evaluated . The results were as follows： the best concentration of the coated antigen and the second antibody were 1 ： 1000 and 1 ： 6000, respectively, the critical value was 0. 131-0. 145. The established ELISA method was much sensitive than IHA. The antibody, which titer was 2^7 tested by IHA, still shows positive detected by ELISA method after 2^14 times diluted. This method is simple, accurate and high specificity with good repeatability.

关 键 词：猪瘟 卵黄抗体 间接ELISA 

分 类 号：S852.3[农业科学—基础兽医学]