Background : Poor adherence is a critical problem in managing Human Immunodeficiency Virus (HIV ) -infected patients receiving Antiretroviral Therapy (ART). Evidence of adherence to antiretroviral therapy after initiation of Universal Test and Treat (UTT) strategy was limited in Ethiopia. Hence, this study aimed to compare adherence to antiretroviral therapy before and after the initiation of universal test and treat strategy, including factors affecting adherence among HIV positive adults in Dessie town. Methods: A comparative cross-sectional study was conducted on 594 HIV positive adults selected using consecutive sampling. Interview and patient record review were used to collect data. The data were analyzed using SPSS version 23. Bi-variable and multivariable logistic regression model were used to identify factors associated with ART adherence. Adjusted Odds Ratio (AOR) with 95% Confidence Interval (CI) was used as a measure of association. Statistical significance was declared at a P - value less than 0.05. Result : The overall proportion of ART adherence using Morisky scale and self-reports were 52.3%, 95% CI: (48.4%, 56.2%) and 95%, 95% CI: (93.5%, 96.8%) respectively. Absence of depression (AOR =3.87, 95% CI: (1.96, 7.64)), eating three or more meals per day (AOR =2.65, 95% CI: (1.08, 6.49)) and absence of concomitant illness (AOR =0.42, 95% CI: (0.23, 0.76)) were significantly associated with better ART adherence. Conclusion: The overall proportion of ART adherence measured by Morisky scale was very low while the proportion of ART adherence measured by self-report was high and consistent with the current World Health Organization (WHO) recommendation. Adherence to ART was not affected by the introduction of the UTT strategy in HIV treatment and care program. Depression, meal frequency and concomitant illness were factors associated with ART adherence. Efforts should be made to improve adherence through tailored interventions to overcome factors linked with poor adherence. Key words: Antiretroviral therapy, Adherence, HIV/AIDS, Ethiopia