中国兽医科学2026，Vol.56Issue(1)：56-62,7.DOI:10.16656/j.issn.1673-4696.2025.0235
牛病毒性腹泻病毒单克隆抗体的制备及抗原捕获ELISA检测方法的建立
Preparation of monoclonal antibodies to bovine viral diarrhea virus and establishment of antigen-capture ELISA detection method
摘要
Abstract
To prepare monoclonal antibodies(McAb)against bovine viral diarrhea virus(BVDV)and establish an ant i gen-capture ELISA detect ion method,BALB/c mice were immuni zed with a truncated BVDV Erns protein expressed in E.coli.After three immunizations,cell fusion and hybridoma screening were performed to select high titer McAb.The BVDV antigen-capture ELISA was established through condition optimization.Its specificity,sensitivity,and reproducibility were validated and compared with a commercial IDEXX kit and RT-PCR for clinical sample fitness.The results showed that two higher titer IgG1 type MAb hybridoma clones(2B3 and 2B10)were obtained,both containing κ light chains.The higher titer 2B10 McAb was chosen as the detection antibody,paired with a rabbit derived BVDV polyclonal anti-body as the capture antibody.The developed BVDV antigen capture ELISA had no cross reactivity with other common bovine pathogens,including bovine rotavirus,bovine coronavirus,bovine parainfluenza virus type 3,bovine respiratory syncytial virus,and Mycoplasma bovis.It had a detection limit of 102.53 TCID50 for BVDV viral particles.Intra and inter assay coefficients of variation were below 5％,and coinci-dence rates with the IDEXX kit and RT-PCR were 91.76％—93.33％and 94.12％—96.67％,respectively.In conclusion,the 2B10 based BVDV antigen-capture ELISA developed has good specificity,sensitivity,and reproducibility,providing a useful technical means for BVDV detection,prevention,and control.关键词
牛病毒性腹泻病毒/Erns蛋白/单克隆抗体/ELISAKey words
bovine viral diarrhea virus/Erns protein/monoclonal antibody/ELISA分类
农业科技引用本文复制引用
赵嘉豪,李树凡,薛凤,王君,张灿,刘文华,徐守振..牛病毒性腹泻病毒单克隆抗体的制备及抗原捕获ELISA检测方法的建立[J].中国兽医科学,2026,56(1):56-62,7.基金项目
国家重点研发计划项目(2018YFD0501403) （2018YFD0501403）
