中国动物传染病学报2024，Vol.32Issue(6)：57-62,6.
赤羽病病毒N蛋白阻断ELISA抗体检测方法的建立及应用
Development of a Blocking ELISA for Detecting the Antibodies Against Akabane Virus N Protein
摘要
Abstract
To develop a blocking ELISA method for detecting antibodies of Akabane virus(AKAV)N protein,the purified recombinant N protein was used as the coating antigen and HRP-labeled monoclonal antibody(mAb)1C5 was used as the blocking antibody.After optimization,a blocking ELISA assay based on AKAV N protein was developed for detecting AKAV antibodies.The optimal blocking ELISA was used to test 67 negative serum samples and the blocking rate(PI)was calculated to determine the cut-offvalue and criterion.A serum sample tested was defined as AKAV antibody positive at PI>53.1%,negative at PI<46.8%and suspicious at PI 46.8%-53.1%.The blocking ELISA had no cross reaction with Bovine viral diarrhea virus(BVDV),Lumpy skin disease virus(LSDV)and Bluetongue virus(BTV),indicating its specificity.In addition,the 79 clinical serum samples were tested using this blocking ELISA and a commercial ID screen AKAV antibody detection kit and their coincidence was 91.1%.The results showed that the blocking ELISA developed here could be used for the detection of AKAV serum antibodies in domestic clinical practice and had very important reference significance for diagnosis of AKAV.关键词
赤羽病病毒/单克隆抗体/阻断ELISA/蛋白表达Key words
Akabane virus/monoclonal antibody/blocking ELISA/protein expression分类
农业科技引用本文复制引用
陈冬杰,魏方,李昊轩,王晶晶,许晓琳,吴绍强..赤羽病病毒N蛋白阻断ELISA抗体检测方法的建立及应用[J].中国动物传染病学报,2024,32(6):57-62,6.基金项目
基本科研业务费(2024JK003) （2024JK003）
国家重点研发计划项目(2022YFD1802000) （2022YFD1802000）
