Kaffir lime (Citrus hystrix DC) is citrus plant of the family Rutaceae that was used as an antibacterial. This plant is native to Indonesia, which contains flavonoids, tannins, saponins, alkaloids, and essential oil. This study aims to determine the ability of antibacterial extract lime leaves against Staphylococcus aureus and to find out the concentration that is most effevtive in inhibiting the bacteria Staphylococcus aureus. The research used lime leaves. By drying the leaves at the oven at temperature 45˚C, pulverizing by mesh 40, and extraction by maceration for 5 days with ethanol 96%. This study used a completely randomized design with treatments in variation concentrations of 5%, 10%, 15%, and 20% concentration were then used to test the antibacterial activity with disc diffusion. Inhibition zone formed from extracts with concentration of 5% (6,7 mm) 10% (6,8 mm) 15% (7,3 mm) and 20% (8,3 mm). The results of the data analysis showed that the administration of lime leaves extract affected the growth of Staphylococcus aureus, where the value of sig <0,05 obtained a value of sig 0,000. Based on the research, among the others the concentration of 20% was that the most effective in inhibiting the growth of Staphylococcus aureus that was 8,3 mm. Kaffir lime (Citrus hystrix DC.) leaf extract has antibacterial activity against the Staphylococcus aureus. Concentration extract 20% is an effective concentration to inhibit bacterially