动物医学进展2016，Vol.37Issue(11)：19-24,6.
牛副流感病毒3型抗体间接 ELISA 检测方法的建立与初步应用
Establishment and Preliminary Application of an Indirect ELISA for Detecting Antibodies against Bovine Parainfluenza Virus Type 3
摘要
Abstract
In order to establish an indirect ELISA (iELISA)for detecting antibodies against bovine parain-fluenza virus type 3 (BPIV3),the truncated NP-HN gene was cloned and pET28a(+)-NP-HN vector of prokaryotic expression was constructed.We optimized the ELISA reaction conditions using the NP-HN re-combinant protein as coating antigen,and established an iELISA method to specifically detect the positive serum of BPIV3.Further the iELISA method was compared with virus neutralization test and imported ELISA kits,respectively.Finally,270 clinical serum samples were tested.The results showed that the re-combinant 44 ku protein could highly expressed in the form of soluble and inclusion bodies in E.coli,which had good antigenic specificity and reactionogenicity,and had no cross reaction with positive serum of bovine viral diarrhea virus,infectious bovine rhinotracheitis virus and other respiratory pathogens.The coefficients of variation for intra and inter-assay were lower than 8%.The total coincidence rate with virus neutraliza-tion test and imported ELISA kit were 96.67% and 98.89%.The total positive rate of 270 clinical serum samples from Shandong,Liaoning,and Tianjin was 82.59% (223/270).These results demonstrated that BPIV3 indirect ELISA method was specific and sensitive,and can be used for epidemiological investigation and antibody detection for BPIV3.关键词
牛副流感病毒 3 型/NP-HN 基因/原核表达/酶联免疫吸附试验/抗体检测Key words
Bovine parainfluenza virus type 3/NP-HN/prokaryotic expression/ELISA/antibody detection分类
农业科技引用本文复制引用
杨建乐,赵贵民,侯佩莉,王洪梅,李杰,何洪彬..牛副流感病毒3型抗体间接 ELISA 检测方法的建立与初步应用[J].动物医学进展,2016,37(11):19-24,6.基金项目
现代农业（奶牛）产业技术体系科学家岗位项目（Cars-37）.山东省自然科学基金培育项目（ZR2015PC007）.山东省农业科学院科技创新重点项目（2014CXZ08）子课题（2014CXZ08-3）.山东省农业科学院重大成果培育项目 （）
