Staphylococcus aureus (S. aureus) has been important human pathogen responsible for wide spectrum of infection ranging from mild to severe life threatening infection. The present study was carried out to assess the prevalence and susceptibility patterns of S. aureus nasal colonization among health care workers (HCWs). This study was done at the Department of Microbiology, Bharati Vidyapeeth Deemed University Medical College and Hospital, Sangli, Maharashtra. A cross sectional study was carried out. Four hundred and fifty (n=450) HCWs were screened for S. aureus by nasal swabbing. S. aureus strains were detected by gram staining, catalase test, slide and tube coagulase test, phosphatase test and antibiotic sensitivity testing by Kirby-Bauer Method. The Z-test was also carried out to detect standard error of difference between two proportions. Of the 450 samples screened 102 (23%) strains of S. aureus were isolated. The overall carriage rate of S. aureus in our study was 23% with the highest rate being seen among the nursing staff 42(28%) and other categories of HCWs i.e. Doctors and Housekeeping staff carriage rate was same 30 (20%) but it was less as compared to the nursing staff. A high rate of resistance (>50%) was found against ampicillin, erythromycin and penicillin. The isolates of S. aureus were more susceptible (>80%) to gentamycin, cefoxitin, vancomycin, cephalexin and tetracyclin. Our study showed that nursing staff were the potential colonisers of S. aureus when compared to other clinical staff. This study documents the prevalence and colonization of S. aureus among HCWs at our institute.