中国兽医科学2018，Vol.48Issue(6)：712-716,5.DOI:10.16656/j.issn.1673-4696.2018.0109
猪源伪狂犬病病毒gB蛋白抗体阻断ELISA的建立与应用
Development of a blocking ELISA for the detection of gBprotein antibodies against pseudorabies virus in swine
摘要
Abstract
In order to develop a blocking ELISA for the detection of the antibodies against pseudorabies virus(PRV) gB protein by using recombinant gB protein of the strong virulent strain PRV ZJ01 and the HRP-lablled monoclonal antibodies against gB protein, an ELISA reaction conditions were optimized. The concentration of the coating antigen was 0. 5 g/ mL, and the serum samples were diluted at 1 ∶ 1 and incubated for 1 h at 37 ℃, and Mab-HRP(1 ∶ 15 000 dilution) was incubated for 0. 5 h at 37 ℃. The criteria of the ELISA results were that the samples were considered to be positive if it's percentage inhibition(PI) was greater than or equal 28. 67％, that the samples were rated to be negative if it's Piwas less than or equal 18. 27％, and that the samples were considered inconclusive if it's Piwas between 18. 27％ and 28. 67％. The ELISA method was proved to have no cross-reaction with positive sera with PRRSV, PCV2, CSFV and FMDV, respectively. The intra-assay and inter-assay variation coefficients were both less than 10％. The comparison experiments showed that the specificity and accuracy of ELISA were 80. 9％ and 96. 4％, re spectively, and the rate of coincidence was 85. 1％ compared with the commercial IDEXX PRVgBantibody test kit. The detection of 535 clinical serumsamples frompiglets showed the positive rate for PRV antibody reached up to 61. 87％. In conclusion, this blocking ELISA was specific, simple and fast, which lay a foundation for the establishment of standardized testing kit for PRV.关键词
猪/伪狂犬病病毒/gB蛋白/阻断ELISAKey words
porcine/pseudorabies virus/gBprotion/blocking ELISA分类
农业科技引用本文复制引用
孙海凤,徐旋,董静,孙涛,白娟,姜平..猪源伪狂犬病病毒gB蛋白抗体阻断ELISA的建立与应用[J].中国兽医科学,2018,48(6):712-716,5.基金项目
江苏省农业科技自主创新资金项目[CX(15)100604] （15）
国家生猪产业技术体系专项(CARS-36) （CARS-36）
国家重点研发计划项目(2016YFD0500105) （2016YFD0500105）
