Background: Healthcare workers (HCWs) can act as asymptomatic carrier in transmitting Methicillin Resistant Staphylococcus aureus (MRSA). Availability of epidemiological and antibiotic susceptibility data is necessary to limit the spread of HCWs-associated MRSA infections, and to help physicians in choosing the appropriate empirical antibiotic for management of such infections. Objective: to assess nasal carriage and antibiogram of MRSA in healthcare workers from Southern Jordan. Methods: a total of 276 nasal swabs were randomly collected from the HCWs. MRSA was identified by culture, biochemical and molecular methods. Antibiotic susceptibility was determined by the disc diffusion method. Results: The HCWs-MRSA nasal carriage was 8.7%. There was significant difference for nasal carriage of MRSA by nurse occupation (p value = 0.007), education level of less than a university degree (p value = 0.039) and years of HCW experience (p value = 0.023). No significant difference by age, sex, antibiotic exposure or smoking. Antibiotic resistance to Trimethoprim-Sulfamethoxazole and Tetracycline was detected in 37.5% and 12.5% of all MRSA isolates respectively. No resistance to the other antibiotics used in this study and no multidrug resistance was encountered in all MRSA isolates. Conclusion: MRSA nasal carriage among HCWs in this study was 8.7% with no alarming antibiotic resistance pattern. Nurses, less educated and more experienced HCWs are at increased risk of MRSA nasal carriage. Therefore, we strongly recommend screening and decolonizing positive HCWs who can act as asymptomatic carriers in MRSA transmission cycle. Keywords: Methicillin resistant Staphylococcus aureus, Healthcare workers, Antibiogram