鸭圆环病毒抗体的间接ELISA检测方法的建立  

Development of an indirect ELISA method for detection of antibody to duck circovirus

作　　者：苏小东[1] 曹瑞兵[1] 张羽[1] 刘文娟[1] 陈昊[1] 祝长青[2] 陈溥言[1] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]江苏出入境检验检疫局,江苏南京210001

出　　处：《南京农业大学学报》2011年第5期117-121,共5页Journal of Nanjing Agricultural University

基　　金：南京农业大学SRT计划项目(1004A14)

摘　　要：鸭圆环病毒(DuCV)是新发现的一种感染鸭的最小病毒,可导致鸭淋巴组织损伤。目前尚无检测其抗体的商品化ELISA试剂盒。为了对鸭群进行DuCV血清流行病学调查,本试验用原核表达了全长DuCV Cap蛋白,Western blot检测证明表达产物具有良好的抗原性。以纯化的重组Cap蛋白作为包被抗原,应用30份DuCV抗体阴性血清和16份DuCV抗体阳性血清,建立了检测DuCV抗体的间接ELISA方法,以此方法对从江苏地区收集的部分鸭血清样品进行检测,结果显示DuCV抗体阳性率为26.7%(38/142)。试验证明该方法具有良好的特异性、敏感性和重复性,且方法的变异系数均小于10%。结论:该方法操作简便,可以低成本、大规模地检测鸭圆环病毒抗体。Duck circovirus is a newly emerged virus that was commonly associated with damage of lymphoreticular tissues and it is the smallest virus isolated from duck so far.There is no commercial ELISA kit available for duck circovirus（DuCV）antibody detection.In order to survey the serum epidemiological characteristic of DuCV,an indirect ELISA was developed based on the recombinant full-length Cap protein of DuCV in the present study.The recombinant Cap protein was expressed in E.coli expression system and purified by His Bind affinity chromatography.The recombinant Cap based ELISA was evaluated by examining a panel of 30 DuCV antibody negative and 16 DuCV antibody positive sera.Cross-reactivity assay showed that this assay was DuCV antibody specific.Repeatability tests revealed that the coefficients of variation of positive sera within and between runs were less than 10%.Using the established indirect ELISA,DuCV-specific antibodies were detected in 38（26.7%）of 142 samples obtained from 8 of 12 duck flocks from Jiangsu Province.This ELISA is simpler to produce and perform,suitable for large scale surveys of DuCV infection at low cost.

关 键 词：鸭圆环病毒 重组CAP蛋白 间接ELISA 

分 类 号：S852.2[农业科学—基础兽医学]