Abstract: In order to understand the drug resistance phenotype and the carrying of drug resistance genes of cow mastitis-derived Staphylococcus aureus in Shihezi area, in this experiment, bacteria isolation was conducted from milk samples of dairy cows with mastitis in two large-scale dairy farms in Shihezi area. Species-specific primers were used to identify the isolates; the detection of the drug-resistant phenotypes and the drug-resistant genes of the obtained Staphylococcus aureus were carried out by K-B method, MIC method and PCR method. The results showed that 74 isolates of Staphylococcus aureus were isolated from 138 milk samples of dairy cows with mastitis, which were positive cocci by microscopic examination, and identified as Staphylococcus aureus by PCR amplification, with an isolation rate of 53.62%（74/138）. All isolates had drug resistance in varyous degrees. The drug resistance rates of vancomycin, methicillin, sulfamethoxazole, lincomycin, penicillin and enrofloxacin were 97.30%, 82.43%, 79.73%, 71.62%, 67.57% and 67.57%, respectively. The drug resistance rates of the other nine drugs were 16.22%-59.46%. The sensitivity to the two drugs gentamicin and doxycycline was 52.70% and 74.32% respectively. 90.54% of the isolates were resistant to three or more antibiotics, and 56.75% of the isolates were resistant to nine or more antibiotics; one isolate was resistant to all test antibiotics. Seven resistance genes, including blaZ, linA, ermB, aacA-aphD, aac6-aph2, Aph（3′）-a and tetM, were detected in the 74 isolates. Among them, the detection rate of aminoglycoside resistance gene aacA-aphD was the highest of 8.10%, and the detection rates of other drug resistance genes were 2.70%-4.05%. The results suggested that the drug resistance of cow mastitis-derived Staphylococcus aureus existed widespread in Shihezi area, Xinjiang Uygur Autonomous Region, and the phenomenon of multiple drug resistance was serious. A recommendation was given that the scientific clinical treatment of dairy cow mastitis should be based on the results of drug sensitivity test.