ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Infectious Agents and Disease
This article is part of the Research TopicRapid and Efficient Analytical Technologies for Pathogen DetectionView all 23 articles
Development of a nucleocapsid protein-based competitive ELISA for the detection of porcine deltacoronavirus antibodies
Provisionally accepted
- 1Yan'an Vocational & Technical College, Yan'an, China
- 2Northwest A&F University, Xianyang, China
- 3Animal Disease Control Center of Yan'an, Yan'an, China
- 4Animal Husbandry and Veterinary Service Center of Baota District, Yan'an, China
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Porcine deltacoronavirus (PDCoV) is a significant threat to the global swine industry, causing severe diarrhea in piglets and exhibiting a broad host range with zoonotic potential. The nucleocapsid (N) protein of PDCoV is highly conserved and immunogenic, making it an ideal target for diagnostic development. However, existing enzyme-linked immunosorbent assays (ELISAs) typically rely on enzyme-labeled secondary antibodies, complicating the experimental procedure. In this study, a monoclonal antibody (mAb 3C6) against the PDCoV N protein was developed and utilized to establish a competitive ELISA (cELISA). The recombinant N protein (~40 kDa) was successfully expressed and purified. Epitope mapping identified the linear B-cell epitope recognized by mAb 3C6 as 121-HQLLPLRFPTGDGPA-135. Using 60 PDCoV-negative serum samples, the cut-off value for the cELISA was determined to be 44.3%. Validation with 175 field swine serum samples demonstrated a diagnostic sensitivity of 88.5%, specificity of 92.6%, and an overall agreement of 92.0%. The cELISA showed analytical sensitivity twice that of an indirect immunofluorescence assay (IFA) and detected PDCoV-specific antibodies as early as 7 days post-infection. No cross-reactivity with other common swine pathogens was observed. The coefficients of variation for intra-and inter-assay reproducibility ranged from 4.39% to 9.20% and 4.20% to 7.97%, respectively. In conclusion, the developed N protein-based cELISA represents a reliable and efficient tool for the serological detection of PDCoV.
Keywords: porcine deltacoronavirus, Competitive ELISA, Monoclonal antibody, N protein, Antibodies
Received: 06 Aug 2025; Accepted: 14 Nov 2025.
Copyright: © 2025 Shuizhong, Yu, Maiping, Penggang, Xingming and Ying. 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: Wang Ying, wangying.052@163.com
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