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ORIGINAL RESEARCH article

Front. Vet. Sci.
Sec. Veterinary Experimental and Diagnostic Pathology
Volume 12 - 2025 | doi: 10.3389/fvets.2025.1530870
This article is part of the Research Topic Innovative Approaches in Veterinary Pathology: Diagnostics, Therapeutics, and Zoonotic Threats View all 4 articles

Establishment of an indirect ELISA method for detecting bovine coronavirus antibodies based on N protein

Provisionally accepted
Qiang Liu Qiang Liu 1Xiaoxia Niu Xiaoxia Niu 1Lingling Jiang Lingling Jiang 1Gang Zhang Gang Zhang 1Pu Wang Pu Wang 1Sinong Zhang Sinong Zhang 1Weifeng Gao Weifeng Gao 1Guo Huichen Guo Huichen 2*Yujiong Wang Yujiong Wang 1*Yong Li Yong Li 1*
  • 1 Ningxia University, Yinchuan, China
  • 2 State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute (CAAS), Lanzhou, Gansu Province, China

The final, formatted version of the article will be published soon.

    Bovine Coronavirus (BCoV) is a significant pathogen responsible for neonatal calf diarrhea, winter dysentery in adult cattle, and bovine respiratory diseases. Infection with the virus can result in hemorrhagic diarrhea, decreased milk production, and potentially fatal outcomes in cattle, leading to considerable economic repercussions for the cattle industry. Efficient management of BCoV relies on swift and precise detection techniques. CHO cells were utilized to express a secreted recombinant nucleocapsid protein (N), whereby rabbit polyclonal antibodies (pAb) were generated through immunization. An indirect enzyme-linked immunosorbent assay (iELISA) based on N protein was established for the detection of BCoV antibodies. Reaction conditions were optimized using a checkerboard approach, with the optimal antigen concentration at 1.25 μg/mL and the optimal antibody dilution at 1:200, the cutoff value distinguishing negative and positive serum samples was 0.986. The sensitivity test indicated that this rabbit pAb had a maximum dilution of 2 18 within the assay range, did not cross-react with BHV-1, BVDV, BRV, and BRSV positive serum samples, and shown great specificity. The developed iELISA method and commercial kit were used to test 58 bovine serum samples, and the concordance rate was 94.83%. In summary, we have developed a cost-efficient and precise iELISA method based on N protein that serves as a useful diagnostic tool for BCoV in clinical samples and epidemiological research.

    Keywords: Bovine coronavirus, N protein, Eukaryotic expression, Indirect ELISA, Antibody detection

    Received: 19 Nov 2024; Accepted: 23 Jan 2025.

    Copyright: © 2025 Liu, Niu, Jiang, Zhang, Wang, Zhang, Gao, Huichen, Wang and Li. 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:
    Guo Huichen, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute (CAAS), Lanzhou, 730046, Gansu Province, China
    Yujiong Wang, Ningxia University, Yinchuan, China
    Yong Li, Ningxia University, Yinchuan, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.