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Background <p>Senecavirus A (SVA) causes considerable economic losses in the swine industry, particularly among piglets. However, effective serological diagnostic assays for SVA are still lacking. A testing platform based on nanobody-horseradish peroxidase (Nb-HRP) fusion protein-based competitive ELISA (cELISA) for the detection of antibodies against various animal viruses is available. In this study, a rapid and reliable cELISA was established using this platform for specific detection of antibodies against SVA.</p> Results <p>Two anti-SVA nanobodies were selected from an immunized Bactrian camel using phage display technology. The Nb84-HRP fusion protein was expressed and used as a probe during the process. The assay exhibited a cut-off value of 19.4% and showed no cross-reactivity with antibodies against other swine viruses, demonstrating high specificity. The intra- and inter-assay coefficients of variation ranged from 1.44% to 3.27% and 1.36% to 8.71%, respectively. Evaluation using clinical swine serum samples showed a higher coincidence rate between the cELISA and Western blot (99%, kappa value = 0.97) than that between the cELISA and a commercial ELISA kit (84%, kappa value = 0.35).</p> Conclusions <p>These results indicate that the cELISA developed in this study is a rapid, cost-effective, and reliable method for serological detection and surveillance of SVA infection.</p>

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Development of a novel nanobody-based competitive ELISA for rapid detection of antibodies against Senecavirus A

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月29日

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  • Longzhi Nie,
  • Tianxiang Chen,
  • Yunpeng Shi,
  • Bing Wu,
  • Chao Xiong,
  • Qin Zhao,
  • Yuchen Nan,
  • Yiyang Chen,
  • Baoyuan Liu

摘要

Background

Senecavirus A (SVA) causes considerable economic losses in the swine industry, particularly among piglets. However, effective serological diagnostic assays for SVA are still lacking. A testing platform based on nanobody-horseradish peroxidase (Nb-HRP) fusion protein-based competitive ELISA (cELISA) for the detection of antibodies against various animal viruses is available. In this study, a rapid and reliable cELISA was established using this platform for specific detection of antibodies against SVA.

Results

Two anti-SVA nanobodies were selected from an immunized Bactrian camel using phage display technology. The Nb84-HRP fusion protein was expressed and used as a probe during the process. The assay exhibited a cut-off value of 19.4% and showed no cross-reactivity with antibodies against other swine viruses, demonstrating high specificity. The intra- and inter-assay coefficients of variation ranged from 1.44% to 3.27% and 1.36% to 8.71%, respectively. Evaluation using clinical swine serum samples showed a higher coincidence rate between the cELISA and Western blot (99%, kappa value = 0.97) than that between the cELISA and a commercial ELISA kit (84%, kappa value = 0.35).

Conclusions

These results indicate that the cELISA developed in this study is a rapid, cost-effective, and reliable method for serological detection and surveillance of SVA infection.