猪脑心肌炎病毒抗体间接ELISA方法的建立及应用  

Development and Application of an Indirect ELISA for Detection of Encephalomyocarditis Virus Antibody in Swine

作　　者：董艳娇[1] 黄金海[2] 王晶钰[1] 赵勇[3] 李琳[2] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]天津大学,300072 [3]天津市动物检疫站

出　　处：《中国动物检疫》2008年第7期30-33,共4页China Animal Health Inspection

基　　金：天津大学青年基金;编号:5110110

摘　　要：应用纯化的EMCV VP1重组蛋白建立了检测EMCV抗体的间接ELISA方法,并确定了间接ELISA的最佳工作条件。结果表明,重组蛋白抗原的最适包被浓度为0.54μg/mL;与间接免疫荧光试验比较,结果表明该方法具有良好的特异性和敏感性。此方法中VP1抗原最佳稀释度为1:100,待检血清的最佳稀释度为1:50,酶标二抗的最佳稀释度为1:800,1%明胶为封闭液,OD450时阴阳性血清的临界值为0.35。对2006年天津7个地区66个猪场295份屠宰猪的血清样本的检测结果显示,天津各地区血清EMCV抗体阳性率在41.67%～93.33%之间,平均84.75%,猪场抗体阳性率在50%～100%,平均93.94%。An indirect ELISA was established by using the purified EMCV recombinant protein VP1 and the optimal reaction conditions were determined. It was showed that the optimal dilution of recombinant VP1 protein for coating plate was 0.54 μg/mL, the dilution of serum sample was 1：50;the working concentration of HRP-labeled rabbit anti-protein IgG was 1：800;the optimal blocking solution was 1% glutin,and the threshold for ELISA was 0.35 at OD450.In the end, The method was applied for detection of EMCV antibodies in 295 serum samples which was collected from 66 pig farms in Tianjin region in 2006.It was found that the sero positive rate of EMCV was averaged 84.59%（41.67% to 93.33%）;The sero positive rate of EMCV from tested farms was from 50% to 100%and the average rate was93.94%. It is necessary to evaluate EMCV infection in slaughter pigs.

关 键 词：猪 脑心肌炎病毒 抗体 间接ELISA 

分 类 号：S858.28[农业科学—临床兽医学] TS207.3[农业科学—兽医学]