Background and objective: Staphylococcus aureus is one of the most important causes of burn infections. Nowadays, resistance to antibiotics is frequently reported due to new mutation in related genes. This study aimed to investigate the in vitro antimicrobial effect of melittin peptide isolated from Persian Honey Bee venom on S. aureus strains isolated from burn infection.Materials and methods: In this study, fifty S. aureus strains were collected from burn infections during 8 months at Shahid Motahari Hospital in Tehran. Melittin was purified from bee venom by reverse phase HPLC using a linear gradient protocol. MIC and MBC Melittin and the Vancomycin determined with the use of Microdilution Broth and CLSI principles.Result: About 22 peaks were detected in chromatogram. Surface area for melittin approximately calculated as 68.9% comparing to the total surface area. According to electrophoresis results the molecular weight of melittin was estimated as 2.8 kDa. MIC50, MIC90, MBC50, MBC90 and also MIC/MBC for Melittin were determined at 1.1, 4.35, 1.1, and 8.7 µg, and 0.86 respectively. MIC50, MIC90, MBC50, MBC90 and also MIC/MBC for Vancomycin was determined at 1, 16, 3, 16 mg, and 0.7 respectively.Conclusions: Melittin inhibited the growth of all clinical strains of S. aureus. According to the results, it was found that inhibition and lethality power of melittin was two folds greater than vancomycin. Good ratio in MIC/MBC for melittin showed that this peptide has the necessary standard index to becoming an antibiotic in future. As melittin induced its antibacterial activity in very small amounts, If melittin will become an antibiotic in future, its complication would be low. Because of powerful effect of melittin, the duration of treatment and also medical expenses would be reduced subsequently.