羊附红细胞体双抗体夹心ELISA诊断方法的建立  

Establishment of Double-antibody Sandwich ELISA for Eperythrozoon ovis

作　　者：张富梅[1] 韩福生[1] 李继连[1] 李子平[1] 

机构地区：[1]河北北方学院动物科技学院,河北张家口075000

出　　处：《中国畜牧兽医》2011年第5期241-243,共3页China Animal Husbandry & Veterinary Medicine

基　　金：河北省科技攻关计划项目(07220410)

摘　　要：为快速准确诊断和检测羊附红细胞体病,及时采取防治措施,建立了检测羊附红细胞体抗原的双抗夹心ELISA诊断方法。选取规模化养殖场羊,镜检附红细胞体红细胞感染率>90%,无菌采取血液,分离羊附红细胞体抗原,制备纯化兔抗羊附红细胞体抗体,应用辣根过氧化物酶标记抗体,进行双抗体夹心ELISA试验。试验结果表明,双抗体夹心ELISA方法的最佳工作条件为:抗体最佳包被量为82.91μg/mL,酶标抗体最适工作浓度为1∶400,抗原最低检出量为7.81μg/mL;而且与支原体、大肠杆菌、葡萄球菌以及牛、猪、兔附红细胞体均不出现交叉反应,表明该方法具有良好的特异性,可用于羊附红细胞体病的诊断和群体检测。To diagnose and detect Eperythrozoonosis on sheep quickly and take preventive measures in time,the double-antibody sandwich ELISA was established.The blood samples of sheep from large-scale farms whose red cell infective rate was over 90% were selected.Eperythrozoon ovis antigen was isolated,rabbit anti-sheep antibody was purified,antibody was labeled with HRP,then double-antibody sandwich ELISA was established.The results showed that the optimal working concentration of IgG was 82.91 μg/mL,while the best enzyme-conjugated antibody concentration was 1∶400,and the detective minimum of antigen was 7.81 μg/mL.The specific test and crossing reaction indicated that the double-antibody sandwich ELISA was a quick,sensitive way adapted to diagnosis and colony check.

关 键 词：羊 附红细胞体 双抗体夹心ELISA 

分 类 号：S854.43[农业科学—临床兽医学]