中国兽医科学2025，Vol.55Issue(12)：1619-1625,7.DOI:10.16656/j.issn.1673-4696.2025.0200
一种A型口蹄疫病毒单抗的设计与制备并应用于cELISA技术
Design and preparation of a serotype A foot-and-mouth disease virus monoclonal antibody and its application in cELISA technology
摘要
Abstract
To address non-specific reactions inherent in traditional polyclonal antibody-based ELISAs for foot-and-mouth disease virus(FMDV),we developed a high-specificity monoclonal antibody(mAb)targeting the VP1 protein of serotype A FMDV topotype SEA-97,prevalent in China and Southeast Asia.The VP1 gene was cloned and expressed recombinantly in a prokaryotic system.A three-tiered screening strategy-initial indirect ELISA,competitive ELISA(cELISA)validation,and serotype specificity con-firmation-yielded a stable hybridoma cell line(10H2-F2)secreting serotype A-specific mAb(isotype:IgG1/κ).An optimized competitive ELISA established with this mAb demonstrated significantly enhanced performance:no cross-reactivity with serotype O FMDV or nine other cloven-hoofed pathogens(e.g.,classical swine fever virus,porcine reproductive and respiratory syndrome virus,porcine parvovirus,porcine circovirus,pseudorabies virus,bluetongue virus,bovine viral diarrhea virus,capripoxvirus,akabane virus;inhibition rate≤22％).A-FMDV-positive serum could be detected at 256-fold dilution-a 2-fold improvement over existing methods.Total concordance(Kappa=0.945 2)with virus neutralization tests was 97.3％.Minimal inhibition rate variation(coefficient of variation for inhibition rate＜0.3％)after 6 months at 4℃/-20℃;excellent inter-assay reproducibility(F=0.2,P＞0.8).This highly specific,sensitive,and stable assay provides a robust tool for precise A-FMDV serosurveillance,vaccine efficacy evaluation,and outbreak monitoring in China and Southeast Asia,addressing a critical diagnostic gap for regionally prevalent strains.关键词
A型口蹄疫病毒/VP1基因/单克隆抗体/cELISA检测方法/血清型特异性Key words
foot-and-mouth disease virus serotype A/VP1 gene/monoclonal antibody/cELISA/sero-logical specificity分类
农业科技引用本文复制引用
谢佳芮,王轶男,寇美玲,苗海生..一种A型口蹄疫病毒单抗的设计与制备并应用于cELISA技术[J].中国兽医科学,2025,55(12):1619-1625,7.基金项目
广西重点研发计划项目(桂科AB25069499) （桂科AB25069499）
云南省创新引导与科技型企业培育计划项目(202304BI090001) （202304BI090001）
云南省"畜禽种质创新及标准化养殖技术推广"专项 （）
云南省科技人才与平台计划项目(202205AF150007) （202205AF150007）
