Abstract: A total of fifty quarter milk samples were collected from dairy cows with mastitis in selected regions of Inner Mongolia（Hohhot and Baotou City）. Standard microbiological methods were used to isolate Staphylococcus aureus（S. aureus） strains and the suspicious isolates were subsequently phenotypically and molecularly identified. Antimicrobial resistance profiles of the isolates were assessed. Eight S. aureus isolates were obtained from the collected samples and the isolating rate was 16.0%; the antimicrobial susceptibility test results revealed that 12.5% of the isolates were resistant to ciprofloxacin, 37.5% of the isolates showed resistance to erythromycin and ampicillin, 75.0% of the isolates exhibited resistance to ofloxacin, and 100% of the isolates were resistant to aztreonam and penicillin. No resistance was detected for rifampicin, compound sulfamethoxazole,chloramphenicol, vancomycin, cefoxitin, gentamicin, tetracycline, clindamycin, oxacillin, tobramycin, linezolid, meropenem, and amoxicillin/clavulanate.