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小反刍兽疫病毒竞争ELISA检测试剂盒生产工艺研究
引用本文:刘晓慧,杨云庆,叶玲玲,祝贺,吕建强,赵文华,颜红,尹尚莲,花群义,杨仕标,张光培,周晓黎,董俊,艾军.小反刍兽疫病毒竞争ELISA检测试剂盒生产工艺研究[J].中国畜牧兽医,2014,41(9):31-34.
作者姓名:刘晓慧  杨云庆  叶玲玲  祝贺  吕建强  赵文华  颜红  尹尚莲  花群义  杨仕标  张光培  周晓黎  董俊  艾军
作者单位:1. 保定出入境检验检疫局, 河北保定 071051;
2. 云南出入境检验检疫局, 云南昆明 650228;
3. 深圳出入境检验检疫局, 广东深圳 518045;
4. 云南省畜牧兽医科学院, 云南昆明 650224
基金项目:国家863项目(2012AA101301);质检公益性行业科研专项(201310093);国家质检总局科技计划项目(2007IK025)。
摘    要:为研究小反刍兽疫病毒(PPRV)竞争ELISA试剂盒的生产工艺,本试验利用昆虫细胞表达的PPRV核蛋白及其单克隆抗体,建立一种特异性高、敏感性强的PPRV竞争ELISA检测方法,并对该方法的各个步骤进行优化,确定该方法的最适条件,从而确定竞争ELISA的操作程序。并用该方法与OIE推荐的PPRV rC-ELISA抗体检测试剂盒同时检测来自西藏、内蒙古共977份临床羊、牛血清样本,结果显示特异性、敏感性、符合率分别达99.89%、93.82%、99.38%。本研究建立的PPRV竞争ELISA方法为该病毒检测试剂盒的研制奠定技术基础,为小反刍兽疫的防控提供科学依据。

关 键 词:小反刍兽疫病毒  N蛋白  单克隆抗体  竞争ELISA  
收稿时间:2014-03-24

Research on the Production Process of Competitive ELISA Test Kit for Peste des Petits Ruminants Virus
LIU Xiao-hui,YANG Yun-qing,YE Ling-ling,ZHU He,LV Jian-qiang,ZHAO Wen-hua,YAN Hong,YIN Shang-lian,HUA Qun-yi,YANG Shi-biao,ZHANG Guang-pei,ZHOU Xiao-li,DONG Jun,AI Jun.Research on the Production Process of Competitive ELISA Test Kit for Peste des Petits Ruminants Virus[J].China Animal Husbandry & Veterinary Medicine,2014,41(9):31-34.
Authors:LIU Xiao-hui  YANG Yun-qing  YE Ling-ling  ZHU He  LV Jian-qiang  ZHAO Wen-hua  YAN Hong  YIN Shang-lian  HUA Qun-yi  YANG Shi-biao  ZHANG Guang-pei  ZHOU Xiao-li  DONG Jun  AI Jun
Institution:1. Baoding Entry-exit Inspection and Quarantine Burea, Baoding 071051, China;
2. Yunnan Entry-exit Inspection and Quarantine Burea, Kunming 650228, China;
3. Shenzhen Entry-exit Inspection and Quarantine Burea, Shenzhen 518045, China;
4. Yunnan Animal Science and Veterinary Institute, Kunming 650224, China
Abstract:In order to research on the production process of competitive ELISA test kit for peste des petits ruminants virus (PPRV), in this test, nucleoprotein (N) protein expressed in insect cell and monoclonal antibody of PPRV were used to establish a high specificity, sensitivity and strong competitive ELISA method for detection of PPRV, and the competitive ELISA procedures were optimized. 977 sheep and cattle serum samples collected from Xizang and Inner Mongolia were assayed with this established competitive ELISA and rC-ELISA antibody kit for PPRV of the OIE at the same time. The detecting results displayed that specificity, sensitivity and accuracy were 99.89%,93.82% and 99.38%. The competitive ELISA method of PPRV established in this assay would provide the basis for research on the production process of ELISA test kit as well as a reference for prevention scientifically.
Keywords:peste des petits ruminants virus (PPRV)  nucleoprotein protein  monoclonal antibody  competitive ELISA  
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