中国畜牧兽医2017，Vol.44Issue(11)：3306-3312,7.DOI:10.16431/j.cnki.1671-7236.2017.11.028
基于PEDV Nsp7蛋白间接ELISA抗体检测方法的建立与应用
Development and Application of a PEDV Nsp7-based Indirect ELISA for Antibody Detection
摘要
Abstract
This study was aimed to establish an indirect ELISA to detect antibodies against different strains of porcine epidemic diarrhea virus (PEDV).Puried nonstructural protein 7(Nsp7) was used as coating antigen,and the indirect ELISA was established by optimizing the ELISA reaction conditions.The results showed that the optimal reaction conditions were as follow:The amount of coating antigen was 0.20 μg/well,and the coating condition was at 37 ℃ incubation for 1 h then 4 ℃ overnight;The working dilution of serum samples and HRP-labelled secondary antibody were 1 ∶ 300 and 1 ∶ 10 000,and the incubation time were 2 and 1.5 h,respectively;TMB substrate incubation time was 15 min.Serum sample was determined as positive when its S/P＞0.1694 and negative when its S/P＜0.1398.The ELISA was specific,reproducible and sensitive.Forty samples of suspected PEDV serum samples were tested by the established ELISA,and the coincidence rate between the ELISA and the commercial kit was 95％.The ELISA established in this study could be used clinically to detect the antibody level of different strains of PEDV,and it also had the potential for early diagnosis of PEDV,providing a basis for the development of effective measures to control PEDV.关键词
猪流行性腹泻病毒(PEDV)/非结构蛋白7(Nsp7)/间接ELISAKey words
porcine epidemic diarrhea virus (PEDV)/nonstructural protein 7 (Nsp7)/indirect ELISA分类
农业科技引用本文复制引用
李红杰,任豪杰,刘延珂,高冬生,赵军..基于PEDV Nsp7蛋白间接ELISA抗体检测方法的建立与应用[J].中国畜牧兽医,2017,44(11):3306-3312,7.基金项目
河南省高校科技创新团队支持计划(14IRTSTHN015) （14IRTSTHN015）
郑州市国际合作交流项目(153PGJHZ203) （153PGJHZ203）
