An Enzyme Linked Immuno-Sorbent Assay (ELISA) for Detecting African Swine Fever Antibodies and Differentiating Infected from Vaccinated Animals (DIVA)

African Swine Fever Virus (ASFV) causes major economic damage to the pork industry. There isno cure and outbreaks require widespread testing and culling of infected animals. ASFV vaccinesare in development but not yet commercially available. Once available a companion diagnostic testwill be needed to differentiate infected from vaccinated animals (DIVA). Serodiagnostic tests suchas enzyme-linked immunosorbent assays (ELISA) can be used to detect antibodies from infectionsand vaccinations. Herein we propose to develop a DIVA ELISA kit for ASFV based on a newvaccine candidate ASFV-G-dI177L (Gladue Lab USDA). Little is known about I177L except thata partial deletion (aa112-177) inhibits ASFV virulence. We used software to predict aa112-117structure solubility and antigenicity and our primary goal is to develop I177L (aa112-177) proteinbased indirect ELISA to sensitively detect ASFV antibodies. We will produce peptides to developan ELISA and validate its performance with naturally and experimentally ASFV infected animalsand ASFV vaccinated animals. We will pair I177L ELISA with a p54/p72 antibody detectionELISA. Currently available p54 and p72 ELISAs have false negative rates depending on theinfection and vaccination status. Thus we aim to develop and optimize a combined antigen p54/p72ELISA which we expect to have greater sensitivity and specificity to detect infections andvaccines. If successful the DIVA ELISA kit (I177L and p54/p72) could provide a criticaldiagnostic tool to accompany the ASFV-G-dI177L vaccine. Following Phase I we will expandvalidation efforts and develop reagents to enhance diagnostic specificity sensitivity and point-of- care applications.