METHODS article
Front. Cell. Infect. Microbiol.
Sec. Molecular Viral Pathogenesis
Volume 15 - 2025 | doi: 10.3389/fcimb.2025.1631027
This article is part of the Research TopicAdvancements in the Understanding, Diagnosis, and Control of Viral Diseases of Cattle, Goats, and SheepView all articles
Expression of BVDV E2 Protein in CHO-S Cells and Development of an Indirect ELISA for Serological Detection
Provisionally accepted
- Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China
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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 interbatch 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.
Keywords: Bovine viral diarrhea virus (BVDV), E2 protein, Indirect ELISA, Serological Diagnosis, Recombinant antigen
Received: 19 May 2025; Accepted: 25 Jun 2025.
Copyright: © 2025 Wang, ZHANG, Hou, Tan and Zhang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Jiahui Wang, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China
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