Establishment of an indirect ELISA antibody detection method based on the stable expression of LSDV P32 protein in CHO-K1 cells
Ruolei Xin, Jiawen Zhang, Yinghui Zhang, Danmei Su, Menglin Li, Dan Liu, Ruizhi Wu, Junjie Zhao, Yuanyuan Zhu, Yebing Liu, Xiaoyun Chen, Yong-Jun Wen, Zhen Zhu
Abstract
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.