中国畜牧兽医2017，Vol.44Issue(6)：1818-1824,7.DOI:10.16431/j.cnki.1671-7236.2017.06.034
猪A型口蹄疫病毒特异性IgA抗体间接ELISA检测方法的建立与初步应用
Development and Primary Application of Indirect ELISA Method for Detecting the IgA Antibody against Foot-and-mouth Disease Virus Type A in Porcine
摘要
Abstract
An indirect ELISA for detecting the IgA antibody against porcine foot-and-mouth disease virus (FMDV) type A was developed by using purified FMDV structural protein VP1 as coating antigen, mouse anti-pig IgA monoclonal antibody as second antibody and HRP-conjugated goat anti-mouse IgG as third antibody.The concentration of coating antigen was optimized as 3.50 μg/mL,the dilution and reaction time of second antibody and third antibody were optimized as 1∶10 000 and 30 min, respectively.There was no cross-reactivity with anti-CSFV, PRRSV and other pathogen specific IgA antibodies.The positive detection rate of FMDV type A infectedsamples was above 90%.The coefficient variation of intra-and inter-assay was ranged from 3.16% to 9.76%.The ELISA method described in this study was proved to be specific and rapid for the detection of FMDV specific IgA antibody.It was potential to be applied for detection the level of FMDV specific IgA and evaluate the effect of mucosal immunity.Besides,it provided a new method for clinical diagnosis of foot-and-mouth disease.关键词
口蹄疫/黏膜免疫/IgA/间接ELISAKey words
foot-and-mouth disease (FMD)/mucosal immunity/IgA/indirect ELISA分类
农业科技引用本文复制引用
王园园,潘丽,吕建亮,张中旺..猪A型口蹄疫病毒特异性IgA抗体间接ELISA检测方法的建立与初步应用[J].中国畜牧兽医,2017,44(6):1818-1824,7.基金项目
"十三五"国家重点研发计划(2016YFD0501503) （2016YFD0501503）
甘肃省科技支撑计划(1604NKCA045-2) （1604NKCA045-2）
国家生猪现代产业技术体系项目(CARS-36-06B) （CARS-36-06B）
