Objective:To determine the frequency of Methicillin resistance staphylococcus aureus (MRSA) and identification of drug susceptibility for MRSA isolates among health care workers (HCWs) of a tertiary care hospital of South Punjab Pakistan.Methods:We included 225 HCWs including laboratory staff, doctors, nurses and paramedical staff in this cross-sectional study. The study was conducted in Nishtar medical university/Hospital Multan. The study duration was July-2016 to April-2017. HCWs having no signs of infections and any other systemic disease were included in this study. We used sterile nasal swab sticks for sample collection for determination of S. aureus prevalence. All these samples were processed in the laboratory for MRSA, methicillin sensitive S. aureus (MSSA) and for antimicrobial sensitivity of S. aureus. Chi-square test was used for comparison of frequency of MRSA and MSSA between different HCWs by assuming p-Value ≤0.05 as significant difference.Results:There were 65.3% (147) female participants and only 34.7% (78) male participants. S. aureus was diagnosed in the nasal flora of 24% (54) participants, out of which 9.3% (21) were MRSA positive and remaining 14.7% (33) were MSSA positive. There was no significant difference in frequency of MRSA and MSSA among different HCWs (p-value 0.79). Amikacin and vancomycin were 100% sensitive for MRSA and MSSA. Clindamycin and ciprofloxacin was 80.9% (17) and 71.4% (15) sensitive for MRSA and 100% and 84.8% (28) for MSSA respectively. While oxacillin and Cefoxitin were 100% (21) resistant for MRSA and sensitive for MSSA.Conclusion:Prevalence of MRSA and MSSA is high among HCWs in Pakistan. Amikacin, vancomycin and clindamycin have high sensitivity for MRSA and can be used for empirical treatment of MRSA in suspected patients.