Objective:To establish a method for detecting antibodies against Porcine Deltacoronavirus(PDCoV).Methods:The nucleocapsid(N)protein of Porcine Deltacoronavirus(PDCoV)was expressed in a prokaryotic system,and the recombinant protein was subsequently identified through techniques such as Western Blot and SDS-PAGE.After obtaining the correctly expressed recombinant protein,it was used as an antigen to optimize the ELISA reaction conditions.The specificity,sensitivity,and reproducibility of the assay were tested,leading to the establishment of an ELISA detection method.Finally,this method was applied to detect clinical samples.Results:The optimized conditions for the ELISA assay included an antigen coating concentration of 2 μg/well,followed by incubation at 37 ℃ for 1 h.Blocking was achieved using 1％BSA at 37 ℃ for 1 h.The optimal dilution concentration for the primary antibody was 1∶1 600,with an incubation time of 1 h.The incubation time for the enzyme-labeled secondary antibody was 60 minutes.Notably,the recombinant protein demonstrated specific reactivity with sera from pigs infected with PDCoV.The established ELISA detection method exhibited excellent sensitivity,specificity,and reproducibility.Conclusion:This study has established an indirect ELISA detection method specifically designed for Porcine Deltacoronavirus(PDCoV),providing an effective approach for detecting antibody levels against PDCoV and supporting the prevention and control of the disease.
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