This study was conducted to evaluate drug resistance profile of Staphylococcus aureus in raw milk, fresh fruit juice and dairy farms settings of Mekelle, Tigray. A cross-sectional study was conducted on the total 258 samples of raw cow milk and fresh fruit juice. Antimicrobial resistance status was also checked for identified S. aureus using various commonly used antimicrobial discs. The overall viable staphylococcal count mean and standard deviation of samples from milk shop, fruit juice and dairy milk were found to be 8.86±107, 7.2 × 107, 8.65±107 cfu/ml, 33.87±106, 6.68±106 and 22.0±106, respectively. Among the total 258 samples, 75 (29.07%) samples were found positive for S. aureus. Proportion of the isolation from milk shop, fruit juice and dairy milk samples were 20 (23.26%), 32 (37.21%) and 23 (26.74%), respectively. Antimicrobial test of the high resistance revealed vancomycin (100%), ampicillin (90.9%). ciprofloxacin (90.9%), ceftaroline (63.6%), penicillin-G (81.8%) and clindamycin (72.7%) whereas they are highly susceptible to some antibiotics like gentamicin (100%), streptomycin (81.8%), norfloxacin (63.6%), chloramphenicol (81.8%), sulfamethoxazole (96%), kanamycin (72.7%), polymixin B (72.7%), erythromycin (72.7%) and tetracycline (81.8); also, some S. aureus also showed multi-drug resistance pattern. The present study, we isolated and determined the drug Resistance profile of S. aruesu in Mekelle city, Northern Ethiopia alarmingly, the S. aureus isolates circulating in the raw cow milk, fresh fruit juice and dairy milk. High level of S. aureus isolation from personnel and equipment besides food samples reveals that the hygiene practice is substandard. Prudent drug use and improved hygienic practice is recommended in the raw cow milk of dairy farms and fresh fruit juice to safeguard the public from the risk of acquiring infections and multiple drug resistance (MDR) pathogenic S.aureus.