(1361-B) Quantitative high throughput screening methods for Rift Valley Fever Virus
Monday, February 5, 2024
2:00 PM – 3:00 PM EST
Location: Exhibit Halls AB
Abstract: Rift Valley Fever Virus (RVFV) is a hemorrhagic fever virus that primarily infects livestock but can also infect humans. It primarily affects the liver and brain and can be transmitted vertically from mother to fetus leading to spontaneous abortions and birth defects. Due to its potential as a zoonotic pandemic pathogen, we sought to develop high throughput assays for screening potential antiviral compounds in a variety of cell types. First, we screened a plethora of cell lines to find suitable candidates for 1536 well miniaturization for cytopathic effect (CPE) and direct antigen staining by immunofluorescence (IFA). We have developed one 1536 well CPE-based assay in Vero using caspase 3/7 glo as the detection reagent with an average Z’ of 0.8 and an average S/B of 8. We also developed several 1536 direct antigen immunofluorescence (IFA)-based assays in human liver cell lines (HepG2 and Huh7). The human cell IFA assays demonstrate Z’ scores of >0.3 with at least 70% of cells infected. We validated these assays with the direct-acting antiviral agent, Ribavirin, and a cell-based antiviral agent, Cycloheximide. We then screened several NCATS libraries including over 3,000 compounds in 7 concentrations each for antiviral activity and toxicity.