Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a significant opportunistic pathogen among human immunodeficiency virus (HIV) patients of Ethiopia. This study aimed at delineating the prevalence, antimicrobial resistance, and biofilm-forming potentials of nasally colonized MRSA among HIV patients in the Arba Minch province of Ethiopia. A cross-sectional study was performed in HIV patients who visit anti-retroviral therapy clinic of the Arba Minch Hospital between February and April 2017. Nasal samples were collected and inspected for Staphylococcus following standard procedures. MRSA was identified using cefoxitin disk and antibiotics sensitivity test was performed as per Kirby-Baur disk diffusion method. The formation of biofilm was inspected using both qualitative and quantitative methods. A total of 307 HIV patients were examined. The overall prevalence of S. aureus was found to be 39.7%. The prevalence of MRSA was 20.8%. The rate of nasal colonization of MRSA was relatively higher among females. In bivariate analysis, MRSA colonization was statistically significant in patients with CD 4 count ≤350 (p value = 0.002) and co-trimoxazole prophylaxis (p value = 0.003). Concomitant resistance to erythromycin, tetracycline, and co-trimoxazole were 48.4%, 45.3%, and 39.0%, respectively. Invariably, all MRSA isolates were 100% sensitive to vancomycin. Of the 64 MRSA isolates, 18.7% were considered as multidrug-resistant. The rate of biofilm formation was 34.3%. The results revealed a high prevalence rate in the nasal colonization of MRSA in HIV patients.