High-Throughput Chemical Screening for Inhibitors of Pathogenicity Island 2.

Anti-virulence therapies are under active investigation as antibiotic alternatives; however, their identification from large-scale chemical libraries poses a unique challenge. The dispensability of virulence factors for growth precludes conventional, optical density-based screening methods. Here, we provide a protocol for high-throughput screening with a cell-based, promoter reporter platform. We describe the use of this method for the identification of anti-SPI-2 inhibitors specific to Typhimurium, which may be modified to investigate other virulence factors. For complete details on the use and execution of this protocol, please refer to Tsai et al. (2020).