猪流行性腹泻病毒抗体间接ELISA检测方法建立

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• 英文论文题名：DEVELOPMENT OF AN INDIRECT ELISA OF PORCINE EPIDEMIC DIARRHEA VIRUS
• 论文作者：孟琼 ; 孔宁 ; 单同领 ; 吴永光 ; 左叶雯 ; 童武 ; 郑浩 ; 李国新 ; 于海 ; 童光志 ; 徐永杰
• 英文论文作者：MENG Qiong ; KONG Nong ; SHAN Tong-ling ; WU Yong-guang ; ZUO Ye-wen ; TONG Wu ; ZHENG Hao ; LI Guo-xin ; YU Hai ; TONG Guang-zhi ; XU Yong-jie ; Xinyang Normal University ; Shanghai Veterinary Research Institute ; CAAS
• 年：2016
• 作者机构：信阳师范学院;中国农科院上海兽医研究所;
• 论文关键词：间接ELISA ; PEDV ; 截短S蛋白
• 英文论文关键词：Indirect ELISA ; PEDV ; truncate S protein
• 会议召开时间：2016-07-20
• 会议录名称：中国畜牧兽医学会兽医公共卫生学分会第五次学术研讨会论文集
• 语种：中文
• 分类号：S858.28
• 学会代码：SYGW
• 会议名称：中国畜牧兽医学会兽医公共卫生学分会第五次学术研讨会
• 会议地点：中国内蒙古通辽
• 主办单位：中国畜牧兽医学会兽医公共卫生学分会
• 学会名称：中国畜牧兽医学会兽医公共卫生学分会
• 页数：9
• 文件大小：657k
• 原文格式：D
• 会议级别：全国

摘要

本研究获得5个猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)S1基因截短基因,原核表达后制备多克隆抗体。ELISA、Western Blot和IFA试验结果表明仅有SE蛋白诱导的多克隆抗体具有免疫活性。因此本试验以SE蛋白为抗原建立间接ELISA抗体检测方法,优化各反应参数并确定临界值为0.155。符合率、重复性及交叉试验结果显示,该ELISA检测方法的符合率为93.7%;批内、批间重复率变异系数均小于10%;具有PEDV特异性。表明建立的ELISA检测方法是特定的、敏感的和可重复的。抗S蛋白的ELISA检测方法的建立为PEDV血清学抗体检测和疫苗评估奠定基础。
Five overlopping fragments covered the gene S1 of porcine epidemic diarrhea virus,were separately expressed and be used to prepare the murine polyclonal antibody.ELISA,Western Blot and IFA assays showed that the polyclonal antibody induced by SE protein has immune activity.Therefore,we developed an indirect enzyme-linked immunosorbent assay(i ELISA) based on the protein SE.The parameters of i ELISA were optimized and the cutoff value determined as 0.155.Coincidence rate,repeatability and Cross-reaction tests showed that the coincidence rate was 93.7%;the coefficients of variation of the intra and inter batch were both less than 10%;and has a good specificity of PEDV.The results suggest this i ELISA is specific,sensitive,and repeatable.Based on i ELISA against S protein,it might be easy to evaluate serological antibody detection and assessment of vaccine.

引文

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