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OA

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.