检测鸡黄病毒血清抗体间接ELISA方法的建立与应用  

DEVELOPMENT OF AN INDIRECT ELISA FOR DETECTION OF SERUM ANTIBODY TO CHICKEN FLAVIVIRUS

作　　者：施少华[1,2] 傅光华[1,2] 程龙飞[1,2] 王建 陈红梅[1,2] 万春和[1,2] 胡思科 宋秀梅 黄瑜[1,2] 

机构地区：[1]福建省农业科学院畜牧兽医研究所,福州350013 [2]福建省畜禽疫病防治工程技术研究中心,福州350013 [3]天津市中升挑战生物工程有限公司,天津300380

出　　处：《中国动物传染病学报》2012年第6期23-28,共6页Chinese Journal of Animal Infectious Diseases

基　　金：福建省省属公益类项目(2011R1025-8)(2011R1025-2);现代农业产业技术体系建设专项资金(CARS-43);福建省"种业创新与产业化工程建设项目"(2011FJZY-9)

摘　　要：本研究以鸡黄病毒FQ-C1株为包被抗原,建立检测鸡黄病毒血清抗体的间接ELISA方法。经优化后确定其最佳工作条件为每孔包被抗原0.57μg,血清以1:640倍稀释,羊抗鸡IgG酶标抗体以1:3200稀释,显色10 min后读取OD450值,P/N值≥2.1的血清为阳性。本实验所建立的诊断方法敏感性高于血清中和试验。对849份来自福建的血清样品进行检测表明,鸡黄病毒的血清阳性率达33.1%,说明鸡黄病毒感染有一定的流行性。An indirect enzyme-linked immuosorbent assay（ELISA） was developed to detect serum antibodies to chicken flavivirus.The ELISA plates were coated with chicken flavivirus FQ-C1 strain.The working parameters were optimized to be 0.57 μg coating antigen in 100 μL per well,test serum dilution at 1：640 and HRP-labeled goat anti-chicken IgG at 1：3200.After 10 minutes of color development with tetramethylbenzidine（TMB） substrate,a threshold of positive/negative signal ratio（P/N） ≥ 2.1 was determined as a positive sample.The ELISA method was shown to be more sensitive than serum neutralization.A total of 849 chicken serum samples collected from Fujian province were tested in indirect ELISA and 284 samples（33.1 %） were positive,indicating that flavivirus was prevalent in chicken flocks in Fujian province.

关 键 词：鸡黄病毒 间接ELISA 血清抗体 

分 类 号：S852.659.6[农业科学—基础兽医学]