Staphylococcus aureus (S.aureus) is one of the predominant causative agents of mastitisrnin dairy herds. Mastitis has a negative impact through economic losses in the dairy sectorrnacross the globe. A cross sectional study design was conducted from November 2018 tornMay 2019 with the objectives to determine prevalence of bovine subclinical mastitisrn,S.aureus and antimicrobial resistance profile and detect antimicrobial resistance andrnvirulence genes in S.aureus isolated from milk with bovine subclinical mastitis in CentralrnEthiopia. A total of 265 lactating exotic zebu cross breed dairy cows were screened forrnbovine sub clinical mastitis using California Mastitis Test (CMT). Different biochemical testsrnand polymerase chain reaction (PCR) were used for the isolation and identification ofrnS.aureus isolates. PCR was performed for detection of antimicrobial resistance genes;rnmethicillin resistance (mecA) and erythromycin resistance (ermA, ermC and mrsA) genesrnand virulence genes (clumping factor A (clfA), enterotoxin A (sea), exfoliative toxin A (eta),rnbeta-hemolysin (hlb), toxic shock syndrome toxin-1(tst-1) and intracellular adhesion Drn(icaD)). Out of 265 lactating dairy cows screened, 49.06% (n=130) of them were positivernfor subclinical mastitis. Out of 130 CMT positive samples,76.92% (n=100) of milk samplesrnwere positive for S.aureus and subjected to nine antimicrobials agents for antimicrobialrnresistance test. A large percent of the isolates were resistant to penicillin (93%) followedrnby ampicillin (87%) and tetracycline (75%). Sixty eight PCR confirmed S.aureus isolatesrnwere obtained from 100 phenotypic positive S.aureus isolates and from this, 17.64% (n=12)rnsamples were contained the mecA gene. No amplification was observed for erythromycinrnresistance (ermA, ermC, and msrA) and virulence (tsst-1, sea, icaD and hlb) genes. Thernfrequency of clfA and eta genes in S.aureus isolates were 25% (n=17) and 22.05% (n=15),rnrespectively. The result obtained in this study suggests that poor management practices arernresponsible for the increase in S.aureus isolation. The high resistance of S.aureus tornantimicrobials contribute in bovine mastitis in these farms may cause health problems in therncommunity consuming raw milk purchased from these farms.