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Abstract <p>Porcine deltacoronavirus (PDCoV) is an emerging enteric coronavirus causing high mortality in neonatal piglets and posing a significant threat to the swine industry. Evidence indicates that PDCoV has cross-species transmission potential and may pose a zoonotic risk, emphasizing the need for reliable serological tools for epidemiological surveillance and vaccine evaluation. Here, we developed a double-antigen sandwich enzyme-linked immunosorbent assay (DAgS-ELISA) based on the PDCoV S1 protein, expressed in CHO cells and used as both coating and HRP-conjugated antigen. The assay reliably detected PDCoV-specific antibodies in sera from pigs, chickens, rabbits, and mice, showing high sensitivity (92.86%) and specificity (99.11%) as determined by receiver operating characteristic curve analysis, with excellent reproducibility. No cross-reactivity was observed with antibodies against other common swine pathogens. Concordance with indirect immunofluorescence assay was 96.18% (kappa = 0.923), and assay results correlated strongly with neutralizing antibody titers (Pearson r = 0.865). Overall, this S1-based DAgS-ELISA provides a sensitive, specific, and cross-species applicable method for PDCoV serological detection, supporting its use for epidemiological surveillance and evaluation of vaccine-induced neutralizing antibodies.</p> Key points <p>• <i>A novel S1-based double-antigen sandwich ELISA was established for PDCoV detection.</i></p> <p>• <i>The assay shows high sensitivity, strong specificity, and broad cross-species applicability.</i></p> <p>• <i>ELISA results strongly correlate with neutralizing antibody titers, aiding vaccine evaluation.</i></p>

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Double-antigen sandwich ELISA based on S1 for cross-species detection of Porcine deltacoronavirus antibodies

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月20日

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  • Yingjie Bai,
  • Ruiming Yu,
  • Liping Zhang,
  • Zhongwang Zhang,
  • Ya Liu,
  • Dongsheng Wang,
  • Peng Zhou,
  • Zhangfu Pei,
  • Huichen Guo,
  • Li Pan,
  • Xinsheng Liu

摘要

Abstract

Porcine deltacoronavirus (PDCoV) is an emerging enteric coronavirus causing high mortality in neonatal piglets and posing a significant threat to the swine industry. Evidence indicates that PDCoV has cross-species transmission potential and may pose a zoonotic risk, emphasizing the need for reliable serological tools for epidemiological surveillance and vaccine evaluation. Here, we developed a double-antigen sandwich enzyme-linked immunosorbent assay (DAgS-ELISA) based on the PDCoV S1 protein, expressed in CHO cells and used as both coating and HRP-conjugated antigen. The assay reliably detected PDCoV-specific antibodies in sera from pigs, chickens, rabbits, and mice, showing high sensitivity (92.86%) and specificity (99.11%) as determined by receiver operating characteristic curve analysis, with excellent reproducibility. No cross-reactivity was observed with antibodies against other common swine pathogens. Concordance with indirect immunofluorescence assay was 96.18% (kappa = 0.923), and assay results correlated strongly with neutralizing antibody titers (Pearson r = 0.865). Overall, this S1-based DAgS-ELISA provides a sensitive, specific, and cross-species applicable method for PDCoV serological detection, supporting its use for epidemiological surveillance and evaluation of vaccine-induced neutralizing antibodies.

Key points

• A novel S1-based double-antigen sandwich ELISA was established for PDCoV detection.

• The assay shows high sensitivity, strong specificity, and broad cross-species applicability.

• ELISA results strongly correlate with neutralizing antibody titers, aiding vaccine evaluation.