SBIR-STTR Award

African Swine Fever Virus (ASFV) causes major economic damage to the pork industry. There is no cure and outbreaks require widespread testing and culling of infected animals. ASFV vaccines are in development but not yet commercially available. Once available a companion diagnostic test will be needed to differentiate infected from vaccinated animals (DIVA). Serodiagnostic tests such as enzyme-linked immunosorbent assays (ELISA) can be used to detect antibodies from infections and vaccinations. Herein we propose to develop a DIVA ELISA kit for ASFV based on a new vaccine candidate ASFV-G-dI177L (Gladue Lab USDA). Little is known about I177L except that a 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) protein based indirect ELISA to sensitively detect ASFV antibodies. We will produce peptides to develop an ELISA and validate its performance with naturally and experimentally ASFV infected animals and ASFV vaccinated animals. We will pair I177L ELISA with a p54/p72 antibody detection ELISA. Currently available p54 and p72 ELISAs have false negative rates depending on the infection 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 and vaccines. 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 expand validation efforts and develop reagents to enhance diagnostic specificity sensitivity and point-of- care applications.