A study was conducted to determine the antimicrobial activities of essential oil from Artemisiae argyi leaves. The sample of the essential oil was analyzed by GC–MS. From 18 compounds representing the oils, Eucalyptole (18.42%), Spathulenol (14.32), 4-Methyl-1-(1-methylethyl)-3-cyclohexen-1-ol (3.10%), 3-Carene (2.64%) appeared as the main components. The screening of antimicrobial activity of the essential oil was evaluated using agar diffusion and broth microdilution methods. Gram-positive bacterial were more sensitive than gram-negative bacterial of the 8 microorganisms, and Staphylococcus aureus ATCC 6538 showed the lowest MIC (0.3125%) and MBC (0.625%). In the disc diffusion assay, Staphylococcus epidermidis ATCC 49134 and Bacillus subtilis ATCC 6633 showed obvious inhibitory activity. Survival curve showed that, 2MIC of Artemisiae argyi essential oil had a lethal effect on Candida albicans within the first 1 h. Results presented here suggest that the essential oil of Artemisiae argyi leaves possesses antimicrobial properties, and provides scientific foundations for exploition of Artemisiae argyi. The sample of the essential oil was analyzed by GC–MS. From 18 compounds represents the oils, Eucalyptole (18.42%), Spathulenol (14.32), 4- Methyl-1-(1-methylethyl)-3-cyclohexen-1-ol (3.10%), 3-Carene (2.64%) appeared as the main components. The screening of antimicrobial activity of the essential oil was evaluated using agar diffusion and Broth microdilution methods. Gram-positive bacterial were more sensitive than gram-negative bacterial of the 8 microorganisms, and Staphylococcus aureus ATCC 6538 with the lowest MIC (0.3125%) and MBC (0.625%). In the disc diffusion assay, Staphylococcus epidermidis ATCC 49134 and Bacillus subtilis ATCC 6633 showed significant inhibitory activity. Survival curve showed that, 2MIC of Artemisiae argyi essential oil had a lethal effect on Candida albicans within the first 1 h. Results presented here suggest that the essential oil of Artemisiae Argyi leaves possesses antimicrobial properties, and provides scientific foundations for exploition of Artemisiae argyi.