Wang Jiahui, Zhang Ao, Hou Zhiwei, Tan Bin, Zhang Shuqin
Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China.
Front Cell Infect Microbiol. 2025 Jul 24;15:1631027. doi: 10.3389/fcimb.2025.1631027. eCollection 2025.
Bovine viral diarrhea virus (BVDV) causes ongoing economic losses to the livestock industry. Monitoring antibodies via enzyme-linked immunosorbent assay (ELISA) is a key tool for ensuring the eradication of BVDV from cattle herds. We developed an indirect ELISA (rE2-iELISA) using CHO-S-expressed recombinant E2 protein, the major immunogenic glycoprotein mediating viral attachment and immune evasion. Optimized assay conditions included: 0.4 μg/well antigen coating, 5% BSA blocking, 1:100 serum dilution, and 1:5000 secondary antibody dilution. The assay demonstrated exclusive specificity for BVDV-1 and BVDV-2 with detection sensitivity to 1:1,500 serum dilution. Validation revealed exceptional diagnostic performance: ROC analysis showed 0.998 AUC (cutoff=0.125), 94.8% concordance with IDEXX ELISA, and the intra- and inter-batch coefficient of variation are both less than 5%. The experimental results indicate that the indirect ELISA detection method based on BVDV rE2 exhibits good sensitivity, specificity, and stability. And a stable serological tool for BVDV surveillance and vaccine efficacy evaluation in cattle populations.
牛病毒性腹泻病毒（BVDV）给畜牧业造成持续的经济损失。通过酶联免疫吸附测定（ELISA）监测抗体是确保牛群根除BVDV的关键工具。我们利用CHO-S表达的重组E2蛋白开发了一种间接ELISA（rE2-iELISA），E2蛋白是介导病毒附着和免疫逃逸的主要免疫原性糖蛋白。优化的检测条件包括：每孔0.4μg抗原包被、5%牛血清白蛋白封闭、血清稀释1:100以及二抗稀释1:5000。该检测方法对BVDV-1和BVDV-2具有特异性，检测灵敏度可达血清稀释1:1500。验证显示出卓越的诊断性能：ROC分析显示曲线下面积（AUC）为0.998（临界值=0.125），与IDEXX ELISA的一致性为94.8%，批内和批间变异系数均小于5%。实验结果表明，基于BVDV rE2的间接ELISA检测方法具有良好的敏感性、特异性和稳定性。并且是用于牛群中BVDV监测和疫苗效力评估的稳定血清学工具。
