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Background Alcohol use disorder in people living with the human immunodeficiency virus is an important public health problem associated with high rates of HIV infection and lower antiretroviral treatment adherence. However, little is known about the prevalence of alcohol use disorders and its impact on ART adherence among people living with HIV/AIDS (PLWHA). Thus, this study aimed to evaluate the magnitude of alcohol use disorder and its impact on medication adherence among PLWHA attending the ART clinic. Methods A hospital based cross sectional study design was conducted at University of Gondar Comprehensive Specialized Hospital (UOGCSH) ART clinic from September 30, 2023 to February 30, 2024. The World Health Organization's Alcohol Use Disorders Identification Tool (AUDIT) was utilized to assess alcohol use disorder. Bivariate and multivariate logistic regression analyses were employed to investigate the relationship between AUDs and other variables. Statistical analysis was conducted using SPSS version 26, with a significance level set at P < 0.05. Results The overall prevalence of AUDs was 38.8%, with hazardous use, harmful use, and alcohol dependence accounting for 24.6%, 6.1%, and 8.1% of the total, respectively. In addition, 67.8% of the participants had medication adherence above 95%. Factors associated with alcohol use disorder were male AOR = 1.52; 95% CI (1.01, 2.30), cigarette smoking AOR = 2.384, 95% CI (1.183, 4.80), educational status AOR = 1.96, 95% CI (1.016, 3.764), and age 45-57 years AOR = 3.05, CI (1.587, 5.871). In present study, compared with social drinkers, harmful drinkers and alcohol dependents showed low medication adherence under 95% (AOR = 4.08 and AOR = 8.89, respectively). Conclusion The study reveals a high prevalence of alcohol use and dependence in PLWHA, underscoring the need for effective detection and intervention strategies particularly for males, cigarette smoker, and age between 54 and 75 years in healthcare and community settings. Alcohol use is strongly associated with poor adherence to antiretroviral therapy (ART). Individuals with hazardous alcohol use are four times more likely to have low adherence, while alcohol-dependent individuals are nearly nine times more likely to have low adherence compared to social drinkers., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)