Background: Healthcare workers (HCWs) are at risk for Methicillin Resistant Staphylococcus aureus (MRSA) nasal carriage, subsequent infection and potential transmission of nosocomial infection. Characterization of MRSA detected in HCWs would give data that can be used for prevention and control measures. Objectives: To detect the prevalence and risk factors for MRSA nasal colonization of HCWs in Mansoura University Children's Hospital (MUCH), then determine if the sources of MRSA isolates are community or hospital acquired by detection of SCCmec IV and Panton-Valentine Leukocidin (PVL) genes. Methodology: Nasal swabs were collected from 100 HCWs and processed to detect Staphylococcus aureus (S. aureus) strains, cefoxitin disc diffusion test of S. aureus isolates was done to select MRSA and Methicillin Sensitive Staphylococcus aureus (MSSA) strains, then all the MRSA isolates were further molecularly characterized. MSSA strains were screened for the presence of mecA gene. A second follow up swab was obtained from positive MRSA carriers in the first swab after three months. Results: The Prevalence of nasal carriage of MRSA among HCWs in MUCH was 25% (twenty-five MRSA) isolates, four of them SCCmec IV and PVL positive CA-MRSA strains (16%) and four MSSA (4%) isolates were all mecA negative, none of them were inducible dormant Methicillin Resistant Staphylococcus aureus (ID-MRSA). Persistent MRSA carriers accounted for 52 % of previously colonized HCWs. Conclusion: a considerable proportion of HCWs harbored CA-MRSA in their nares.