Background: Ever-increasing numbers of infections by methicillin-resistant Staphylococcus aureus (MRSA) strains are being reported in the hospitals. Healthcare workers (HCWs) become colonized with MRSA and act as source as well as vectors for the transmission of MRSA infections to the patients. Aim and Objectives: The aim of this article is to detect the carrier rate of methicillin-sensitive Staphylococcus aureus (MSSA) and MRSA among the HCWs and to determine the antimicrobial susceptibility of the isolates to commonly used antimicrobial agents. Materials and Methods: Nasal swabs from 265 HCWs were collected between October 2018 and April 2019. The swabs were processed using standard laboratory techniques. Results: Of the 265 samples collected from HCWs, 46 (17.35%) showed the growth of S. aureus and among them 38 were found to be MRSA. The overall carrier rate of MRSA was 14.33%. The maximum carriage rate for MRSA was seen among nursing staff and that for MSSA among undergraduate medical students. Overall knowledge and attitude levels of HCWs though were found to be better proper preventive practices were not followed. Most of the MRSA strains were sensitive to vancomycin and linezolid, but more than 50% were resistant to commonly used antimicrobial agents in our hospital. Conclusion: Along with strict enforcement of infection control practices, regular surveillance of HCWs for MRSA colonization is necessary to limit the spread of MRSA by HCWs to susceptible patients.