Abstract: In order to understand the prevalence of the resistance phenotype, resistance genes, and virulence genes of calve-derived Staphylococcus aureus in Inner Mongolia Autonomous Region, in this experiment, 104 calf samples（37 nasal swabs, 13 oral swabs, and 54 fecal samples） were collected for the isolation and purification, biochemical identification, and molecular biological identification of calve-derived Staphylococcus aureus. Drug sensitivity test, drug resistance gene detection and virulence gene detection were carried out on the isolated calve-derived Staphylococcus aureus. The results showed that through isolation, cultivation, and biochemical identification of calf samples, 43 strains of Gram positive cocci were obtained, which were preliminarily determined as calve-derived Staphylococcus aureus, with a separation rate of 41.3%. PCR amplification of the 16S rDNA gene from 43 isolated strains yielded a 1 505 bp target band, which was consistent with the expected results. The isolates were resistant to ampicillin, cefoxitin, compound sulfamethoxazole, kanamycin, and the drug resistance rates were 100%, 90.7%, 81.4% and 60.5% respectively, with the lowest resistance rate to ciprofloxacin, which was 9.3%. Among the resistance genes, the detection rate of blaZ gene was the most, 90.7%; the detection rates of norA and norB genes were the same, both 81.4%; the detection rate of ermA gene was 30.2%; the detection rate of ermB gene was 39.5%; the detection rate of emrC gene was 14.0%; the detection rate of Sul1 gene was 53.5%; the detection rate of carrying 7 resistance genes was 4.7%. The highest detection rate of hla gene in virulence genes was the highest, 58.1%; the detection rate of hlb gene was 27.9%; the detection rate of pvl gene was 18.6%, and the remaining virulence genes were not detected. The results indicated that the resistance of calve-derived Staphylococcus aureus in Inner Mongolia Autonomous Region was severe, carrying multiple resistance genes, as well as hla, hlb, and pvl virulence genes. It was recommended to scientifically and reasonably select antibiotics based on drug sensitivity test results in clinical practice to effectively prevent and control the epidemic.