Staphylococcus aureus is an important pathogen of humans and animals. The aim of this study was to characterize 71 of S. aureus isolates from raw cow milk in southern Xinjiang of China, including molecular typing, distribution of virulence genes, and antimicrobial susceptibility. The isolates belonged to 18 sequence types (STs) (including 11 novel STs) and 6 spa types which were divided into five different clonal complexes (CCs), including CC188, CC352, CC22, CC398, and CC5406. The majority of the strains was grouped into multilocus sequence typing (MLST) CC188 (n = 41), t189 (n = 40), and ST5796 (n = 17). Only 30.9, 12.7, 11.3, and 9.9% of the isolates were resistant to erythromycin, clindamycin/norfloxacin, tetracycline, and gentamicin, respectively. Nine of multidrug resistant (MDR) isolates were observed which was associated with CC398-t2876. The adhesion molecules clfa, clfb, and hlb were most frequently detected with the percentage rate of 98.6% (70/71), 98.6% (70/71), and 90.1% (64/71), respectively. The percentage rates of the staphylococcal enterotoxin genes sea, seb, sec, sed, seg, and sei in S. aureus isolates were 5.6, 19.8, 40.8, 1.4, 49.3, and 30.9%, respectively. The see, seh, and sej genes were not found. This study provides data about the occurrence of S. aureus in raw cow milk, revealing high carriage frequency, drug resistance, and population structure of S. aureus. Furthermore, this study suggests that effective hygienic measures be taken when handling dairy cows, in order to prevent spreading MDR strains to human through direct contact and/or consumption of contaminated food.