Background: Early epidemiological studies indicated that 5% of women were colonizedrnwith Staphylococcus aureus (S. aureus) in their genital tract. Postpartum women had thernhighest colonization rates. Studies, mostly from United States have reported that vaginalrncolonization rates for S. aureus in pregnant women range from 14% to 17.1% and it is thernmajor cause of surgical site infection. It was also indicated that between 25% and 50% ofrnwomen undergone cesarean section develop post surgical site infections because of S. aureus. rnObjective: The aim of this study was to determine the prevalence of genital tract colonizationrnof S. aureus and MRSA among pregnant women at Ayder teaching and referral hospital ofrnMekelle University. rnMethods: A cross sectional study was carried out from December, 2011 and February, 2012rnusing non probable convenient sampling technique for screening vaginal cultures for S.rnaureus obtained from 190 pregnant women (at and greater than 24 weeks of gestation) andrnwere processed for identification of S. aureus including methicillin- resistant strains. rnResults: From 190 pregnant women culture result was available from 184 pregnant womenrnwith culture data in the study and of these 43 (23.4%) were colonized with S. aureus. Out ofrnthe 43 isolates, 12 (27.9%) were MRSA positive. The antimicrobial susceptibility pattern forrn43 S. aureus of the isolates showed 100 % resistant to ampicillin , amoxycillin , and penicillinrnfollowed by high resistance to tetracycline (83.7%), erythromycin (41.9%), oxacillin (27.9%),rnciprofloxacilin (18.6%), gentamicin (4.6%), and to vancomycin (2.3%). Multi-resistance torntwo or more antimicrobial agents was observed in 100% of all tested S. aureus. rnConclusion and recommendation: This study provided data on the vaginal carriage rate ofrnMRSA and initial information on the antibacterial resistance pattern in S. aureus obtainedrnfrom randomly selected pregnant women. In our results, high level of MRSA and multi drugrnresistance was observed. Therefore, we recommend that additional studies with morernepidemiologic tools are needed to further asses the role of S. aureus and MRSA colonizationrnin pregnant women in this study area.