鸭出血症病毒间接ELISA检测方法的建立与应用  

Establishment and Application of an Indirect ELISA for Detecting DHDV

作　　者：朱海侠[1,2] 万春和[1] 黄瑜[1] 程龙飞[1] 施少华[1] 傅光华[1] 陈红梅[1] 

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建省畜禽疫病防治工程技术研究中心,福建福州350013 [2]福建农林大学动物科学学院,福建福州350002

出　　处：《中国畜牧兽医》2013年第2期41-44,共4页China Animal Husbandry & Veterinary Medicine

基　　金：现代农业产业体系建设专项资金(CARS-43);福建省种业创新与产业化工程项目(FJZYCX-9);福建省农科教项目(闽财指(2007)573号);福建省发改委项目(闽发改投资(2008)798号文件)

摘　　要：为了建立快速检测鸭出血症病毒(DHDV)的血清学方法,本试验利用浓缩纯化的DHDV作为包被抗原,建立了检测DHDV血清抗体的间接ELISA方法,并对各种检测条件进行了优化。试验结果表明,抗原最佳稀释浓度为7.06μg/孔;最佳包被条件为37℃1h后,4℃包被过夜;待检血清的最佳稀释倍数为1∶25。在优化条件下,阴阳性临界值判定标准为0.44。建立的ELISA方法对鸭瘟病毒、鸭病毒性肝炎病毒、雏番鸭细小病毒和番鸭呼肠孤病毒阳性血清均无交叉反应,结果表明该方法具有良好的特异性。批内和批间重复性试验的最大变异系数分别为0.0221、0.0032,显示该方法具有很好的稳定性,与血清中和试验的符合率为100%。该方法快速、简单、特异性好、重复性好,可用于大批量监测鸭群DHDV血清抗体感染情况。An indirect ELISA was established to detect antibody against duck hemorrhagic disease virus （DHDV） using pu- rified DHDV as coating antigen. The reaction conditions were optimized, including 7.06 ~g/well coating antigen of the virus, 1 ： 25 dilution of testing serum and 1 ： 500 dilution of HRP conjugated anti-duck IgG with cutoff-value of 0.44 （D450 nm）. The specific tests showed that there were no cross-reaction to the anti serum against duck plague virus, duck hepatitis virus, Mus covy duck parvovirus, Muscovy duck reovirus, which showed that the ELISA was specific to anti serum against DHDV. The intro-and inter-assay indicated that the coefficient of maximum variation was 0. 0221 and 0. 0032, respectively, which showed that the method had good stability. And the result was the same as the neutralization test. The method was proved to be rapid, specific and repetitive by detecting different serum samples from immune ducks, which might be used as a serodiagnosis of DHDV.

关 键 词：鸭出血症病毒 间接ELISA 抗体 

分 类 号：S852.65[农业科学—基础兽医学]