The wound is a suitable site for the incidence of resistance infection. Thus, the research for the finding of effective drugs against this infection is necessary. The study was planned to determine the minimum inhibitory (MIC) and minimum bactericidal concentrations (MBC) of propolis ethanolic extract against three pathogens isolated from patients with wound infection. For the aim of the present study, 26 bacteria (10 S. aureus, 10 P. aeruginosa, 6 K. pneumoniae) isolated from wound infection were assessed for their sensitivity to 12.5, 25, 50,100, and 200 mg/ml concentrations of propolis extract using broth dilution method. The majority (70%) of S. aureus isolates were showed a MIC and MBC at 100 and 200 mg/ml concentrations of propolis extract, respectively. In contrast, 50% of P. aeruginosa isolates reported MBC at 200 mg/ml. Notably, 66.7% of K. Pneumoniae isolates were resistance to the used concentrations of propolis extract. Accordingly, this study underlined the antimicrobial activity of Propolis ethanol extract against S. aureus, P. aeruginosa, and K. pneumoniae isolates. Further deep and confirmatory studies are important.