Resistant bacterial infections are a major public health problem, requiring the search for new therapeutic alternatives. This study aims to evaluate in vitro the antibacterial activity of essential oil extracted from Drypetes gilgiana leaves on strains of Escherichia coli and Staphylococcus aureus isolated in Kisangani (DR Congo). The essential oil was obtained by hydrodistillation and then characterized organoleptically and physicochemically. Its antibacterial activity was analyzed using the agar diffusion method and by determining the minimum inhibitory concentration (MIC). The extraction yield was low (0.086%), but the essential oil had physicochemical and organoleptic characteristics favorable to its biological activity. The oil showed very strong antibacterial activity against E. coli (inhibition diameter 78 mm) and moderate activity against S. aureus (9 mm). The minimum inhibitory concentration (MIC) confirmed a higher sensitivity for E. coli (40%) than for S. aureus (80%). These results suggest that D. gilgiana essential oil could be a promising source of antimicrobial agents, warranting future studies to isolate the active compounds and evaluate their mechanism of action.