Development and validation of indirect ELISA for antibody detection against different protein antigens of porcine epidemic diarrhea virus in the colostrum and milk of sows

The objectives of this study are to develop and optimize indirect ELISA based on three coating antigens of porcine epidemic diarrhea virus (PEDV), recombinant spike (S12), nucleocapsid (N), and whole viral (WV) proteins, for the detection of IgG and IgA antibodies in colostrum and milk and to evaluate the diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the assay as a diagnostic method. Colostrum (n = 347) and milk (n = 272) samples from sows were employed in this assay. Indirect ELISA based on three coating antigens was assessed by receiver operating characteristic (ROC) curve analysis with a virus neutralization (VN) test as a reference method, and the cutoff value for calculating DSe and DSp was determined. S12-ELISA showed higher DSe and DSp of IgG and IgA detection compared to N- and WV-ELISA in both colostrum and milk samples. Moreover, S12-ELISA showed perfect agreement and a high correlation with the VN test, which was better than the N- and WV-ELISA for both IgG and IgA detection in colostrum and milk. In contrast, N-ELISA showed lower DSe and DSp compared to S12- and WV-ELISA, along with a correlation with VN and substantial agreement with the VN test. Nevertheless, our developed ELISAs have accuracy for repeatability in both inter- and intra-assay variation. Overall, this research demonstrates that S12-ELISA is more suitable than WV- and N-ELISA to detect IgG and IgA antibodies against PEDV from both colostrum and milk samples.