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Getah virus (GETV) is a mosquito-transmitted disease that affects animals, causing fever, aseptic meningitis, and abortion. Its prevalence in China poses risks to both animal health and public well-being. Currently, there is a scarcity of seroepidemiological data on GETV due to the absence of commercial antibody detection kits for pigs. The aim of this study is to develop a rapid, accurate, and sensitive ELISA, providing a reliable tool for GETV seroepidemiology and laying the foundation for future commercial assay development. In this study, we removed specific hydrophobic domains and intracellular structures from E2 proteins and constructed the recombinant plasmid pCold-TF-E2. The recombinant protein was expressed using a prokaryotic expression system, and efficient purification of the rE2 protein was achieved using a nickel affinity column. The purified rE2 protein is suitable for the development of an indirect ELISA (rE2 ELISA). Following the optimization of reaction conditions for the rE2-ELISA, the cut-off value was 0.356. Additionally, the rE2-ELISA method showed a positive rate of 37.1% for IgG antibodies against GETV when testing 986 pig clinical serum samples collected from pigs in Sichuan between May 2022 and September 2022. The rE2-ELISA method displayed a 95.1% overall agreement with VNT, boasting a sensitivity of 98.2% and a specificity of 92.6%. These results indicate that IgG ELISA based on rE2 protein is an efficient and economical method for the detection of GETV antibodies in pigs, facilitating the diagnosis and prevention of GETV. • An indirect ELISA method for detecting GETV antibodies in swine was established. • The soluble recombinant E2 protein acts as the coated antigen. [ABSTRACT FROM AUTHOR]

Porcine deltacoronavirus (PDCoV) is an emerging discovered coronavirus that causes significant losses in the global swine industry. This study aimed to establish an indirect ELISA method for detecting PDCoV antibodies using the truncated gene of PDCoV spike protein (S). The purified S protein was used as the coating antigen for the polyclonal antibody. The conditions were optimized to establish an indirect ELISA detection method for PDCoV based on the S protein, which showed good specificity and no cross-reaction with SVV-VP1, ASFV-P72, GETV-E2, PRV-gE, etc. The method has high repeatability, with coefficients of variation within and between batches less than 10%. Compared with the commercial kit, the positive coincidence rate is 86.40%, the negative coincidence rate is 89.43%, and the total coincidence rate is 91.76%. This ELISA can be used for PDCoV serological investigation and antibody evaluation. It can also lay the foundation for further research and development of PDCoV S protein ELISA antibody detection kit. • This method is an indirect ELISA detection method established for the neutralizing epitope region in the PDCoV S protein. • This method utilizes prepared polyclonal antibodies as the coating antigen. • The detection rate of this method in Sichuan Province is 91.76%. [ABSTRACT FROM AUTHOR]