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Background <p>Lumpy skin disease (LSD), caused by infection with the lumpy skin disease virus (LSDV), is a highly infectious disease that poses a notable challenge to the cattle industry worldwide. To conduct epidemiological monitoring of LSDV infection in cattle and evaluate the immune efficacy of LSDV vaccines, it is essential to develop a rapid, sensitive, and specific ELISA-based antibody detection method.</p> Results <p>We utilized the LSDV P32 protein, stably expressed in a CHO-K1 suspension cell system, as a coating antigen to develop an indirect ELISA for Capripoxvirus (CaPV) antibody detection. This method specifically recognizes CaPV-positive sera without cross-reactivity with sera positive for bovine viral diarrhea virus, bovine rotavirus, infectious bovine rhinotracheitis virus, and Brucella antibodies. The method demonstrated a maximum serum dilution detection capacity of 1:3200, with intra- and inter-assay variation coefficients below 10%. Comparison with a commercially available kit showed an agreement of 95.7%.</p> Conclusion <p>The indirect ELISA antibody detection method established exhibited excellent specificity, sensitivity, and reproducibility, providing a reliable tool for clinical detection and epidemiological surveys of LSDV. This method offers significant potential for the prevention and control of LSD outbreaks.</p>

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Establishment of an indirect ELISA antibody detection method based on the stable expression of LSDV P32 protein in CHO-K1 cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年6月4日

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  • Ruolan Xin,
  • Jiawen Zhang,
  • Yinghui Zhang,
  • Daoming Su,
  • Menglin Li,
  • Dan Liu,
  • Ruizhi Wu,
  • Junjie Zhao,
  • Yuanyuan Zhu,
  • Yebing Liu,
  • Xiaoyun Chen,
  • Yongjun Wen,
  • Zhen Zhu

摘要

Background

Lumpy skin disease (LSD), caused by infection with the lumpy skin disease virus (LSDV), is a highly infectious disease that poses a notable challenge to the cattle industry worldwide. To conduct epidemiological monitoring of LSDV infection in cattle and evaluate the immune efficacy of LSDV vaccines, it is essential to develop a rapid, sensitive, and specific ELISA-based antibody detection method.

Results

We utilized the LSDV P32 protein, stably expressed in a CHO-K1 suspension cell system, as a coating antigen to develop an indirect ELISA for Capripoxvirus (CaPV) antibody detection. This method specifically recognizes CaPV-positive sera without cross-reactivity with sera positive for bovine viral diarrhea virus, bovine rotavirus, infectious bovine rhinotracheitis virus, and Brucella antibodies. The method demonstrated a maximum serum dilution detection capacity of 1:3200, with intra- and inter-assay variation coefficients below 10%. Comparison with a commercially available kit showed an agreement of 95.7%.

Conclusion

The indirect ELISA antibody detection method established exhibited excellent specificity, sensitivity, and reproducibility, providing a reliable tool for clinical detection and epidemiological surveys of LSDV. This method offers significant potential for the prevention and control of LSD outbreaks.