In this study, we investigated the antibacterial and antifungal activities of hydro-methanolic (MeOH), ethyl acetate (EtOAc), and butanolic (n-BuOH) extracts from the leaves of Dianthus sylvestris subsp. aristidis (Batt) Greuter & Burdet against 78 clinical and 6 reference bacterial and fungal strains using disk diffusion to asses inhibition diameters (ID) and broth dilution methods to determine the minimum inhibitory concentration (MIC). The antimicrobial activity varied the extracts and strains, with interesting antibacterial effects against most tested bacterial strains. Inhibition diameters ranging from 10 to 20 mm, and MIC values varied between 31.25 and 1000 µg/mL. Furthermore, significant antifungal effects were observed, especially against Candida albicans, with ID ranging from 10 to 14 mm and MIC values ranging from 31.25 to 1000 µg/mL. All extracts showed growth inhibition percentages up to 100% against Aspergillus niger. These findings suggest that Dianthus sylvestris subsp. aristidis extracts are promising candidates for developing drugs against resistant pathogens.