中国兽医科学2025，Vol.55Issue(8)：1011-1017,7.DOI:10.16656/j.issn.1673-4696.2025.0150
口蹄疫病毒通用型结构蛋白抗体间接ELISA检测方法的建立
Development of a universal indirect ELISA for the detection of structural protein antibodies against foot-and-mouth disease virus
摘要
Abstract
To establish a universal structural protein antibody detect ion method for foot-and-mouth disease virus(FMDV)serotypes O,A,and Asia 1,this study utilized a short peptide from the highly con-served N-terminal region of the structural protein VP2 of FMDV as the antigen.After optimizing the conditions,an indirect ELISA method was developed to detect antibodies against structural proteins of different FMDV serotypes in cattle.The sensitivity,specificity,repeatability,and agreement with the liquid-phase blocking ELISA(LPB-ELISA)of this method were analyzed.The results showed that the opti-mal coating concentration of the short peptide was 2 μg/mL,the best blocking solution was 1％BSA+5％sucrose,the optimal serum dilution ratio was 1∶4,and the enzyme-labeled secondary antibody dilution ratio was 1∶5 000.Using Maximizing Youden's Index as the cutoff value,the sensitivity was determined to be 87.5％,the specificity was determined to be 93％.No cross-reactivity was observed with positive sera for bovine viral diarrhea virus(BVDV)and infectious bovine rhinotracheitis virus(IBRV).Both in-ter-batch and intra-batch coefficients of variation were≤10％.The agreement rate with LPB-ELISA was 92.59％.This method is time-efficient,simple to operate,and capable of detecting antibodies against structural proteins of different FMDV serotypes in cattle serum,providing a universal approach for the diagnosis of FMDV infection and the evaluation of herd immunity status.关键词
口蹄疫病毒/结构蛋白VP2/通用型/间接ELISAKey words
foot-and-mouth disease virus(FMDV)/structural protein VP2/versatile/indirect ELISA分类
农业科技引用本文复制引用
张慧艳,李坤,孙梦阳,邢向川,付元芳,李平花,李冬,卢曾军,曹轶梅..口蹄疫病毒通用型结构蛋白抗体间接ELISA检测方法的建立[J].中国兽医科学,2025,55(8):1011-1017,7.基金项目
甘肃省科技重大专项(23ZDNA007) （23ZDNA007）
中国农业科学院兰州兽医研究所所级重点任务(CAAS-ASTIP-JBGS-20230402) （CAAS-ASTIP-JBGS-20230402）
