Development and validation of a high-content screening assay for inhibitors of enteropathogenic E. coli adhesion

Abstract Enteropathogenic E. coli (EPEC) causes intestinal infections leading to severe diarrhea. EPEC attaches the host cell causing lesions epithelium coupled with effacement of microvilli. In process, actin accumulates into a pedestal-like structure under bacterial microcolonies. We designed an automated fluorescence microscopy-based screening method for discovering compounds capable inhibiting adhesion and virulence using aurodox, type three secretion system (T3SS) inhibitor, as positive control. The assay employs strain (2348/69) expressing fluorescent protein staining monitoring bacteria their pedestals respectively, analyzing these custom image analysis pipeline. allows discovery preventing formation pathogenic rearrangements. These may be interfering virulence-related molecular pathways relevant developing antivirulence leads.