Background : Viral load monitoring is used as an important marker for diagnosing early treatment failure in patients with HIV infection/AIDS. Ethiopia has started targeted viral load monitoring recently. However, factors leading to virological failure are not well understood. The aim of this study is to assess magnitude of virologic failure and associated factors among adult patient on ART at Debre Markos Referral Hospital, North West Ethiopia. Methods : Cross sectional study was conducted on 304 participants who had started first line HAAR. Data were collected from patients’ chart starting from ART commencement and face to face interview using semi structured questionnaire. Viral load from separated plasma were analyzed according to protocols. The collected data were analyzed using SPSS version 20. Binary and multiple logistic regression models were fitted to identify factors associated with virological failure among adult patient on ART and to control confounding effect. The results were presented as odds ratio (OR) with 95% confidence intervals. Independent associations were considered with p<0.05. Results: Magnitude of virological failure was 10.5%. Lower income, (AOR = 3.5, 95% CI = 1.2 - 10.5, P = 0.024), lack of social support (AOR = 2.9, 95% CI = 1.01 – 8.2, P = 0.024), Interruption of ART, (AOR = 3.5, 95% CI = 1.01 – 12.1, P = 0.046), drug adherence (AOR = 3.6, 95% CI = 1.1 – 11.3, P = 0.028), non-working functional status (AOR = 3.5, 95% CI = 1.2 – 9.7, P = 0.018), WHO stage III or IV (AOR = 2.9, 95% CI = 1.01 – 8.0, P = 0.040), CD4 count < 200 cells/ml (AOR = 3.0, 95% CI = 1.1 – 8.0, P = 0.031) and TB co-infection (AOR = 3.7, 95% CI = 1.2 – 11.3, P = 0.018) were significantly associated with virological failure. Conclusions and recommendations : The prevalence of virologic failure was high. Lower income, lack of social support, interruption of ART, drug non-adherence, baseline non-working functional status, WHO stage, CD4 count < 200 cells/ml and TB co-infection Conducting faith healing, TB co infection, WHO stage III and IV, adherence, and income were determinants of virologic failure. Therefore early identification of associated factors and monitoring of virologic failure has to be strengthened to benefit patients to prevent from further complication. Keywords : HIV, virological failure, ART, Ethiopia