Development and evaluation of a monoclonal antibody-based competitive ELISA for detecting porcine deltacoronavirus antibodies

Abstract

Abstract Porcine deltacoronavirus (PDCoV) is an emerging swine enteropathogenic coronavirus that can cause acute diarrhea and vomiting in newborn piglets and poses a potential risk for cross-species transmission. It is necessary to develop an effective serological diagnostic tool for the surveillance of PDCoV infection and vaccine immunity effects. In this study, we developed a monoclonal antibody-based competitive ELISA (cELISA) that selected the purified recombinant PDCoV nucleocapsid (N) protein as the coating antigen to detect PDCoV antibodies. To evaluate the diagnostic performance of the cELISA, 122 swine serum samples (39 positive and 83 negative) were tested and the results were compared with an indirect immunofluorescence assay (IFA) as the reference method. By receiver operating characteristic (ROC) curve analysis, the optimum cutoff value of percent inhibition (PI) was determined to be 26.8%, which showed excellent diagnostic performance, with an area under the curve (AUC) of 0.9919, a diagnostic sensitivity of 97.44% and a diagnostic specificity of 96.34%. Furthermore, there was good agreement between the cELISA and virus neutralization test (VNT) for the detection of PDCoV antibodies, with a coincidence rate of 92.7%, and the κ analysis showed almost perfect agreement (κ = 0.851). Overall, the established cELISA showed good diagnostic performance, including sensitivity, specificity and repeatability, and can be used for diagnostic assistance, evaluating the response to vaccination and assessing swine herd immunity.

Keywords

• Porcine deltacoronavirus (PDCoV)
• Competitive ELISA (cELISA)
• Antibody detection
• Monoclonal antibody
• Nucleocapsid (N) protein