ABSTRACT: BACKGROUND:Cryptococcosis is a global public health important infectious disease. HIV infection is the main risk factor estimated to account for 95% of cases in the middle- and low-income countries and 80% of the cases in high-income countries. OBJECTIVE:The main aim of the study was to determine the prevalence and associated risk factors of Cryptococcal antigenemia (CrAg) among HIV/AIDS Patients on second-line ART Therapy at Ambo General Hospital and Nekemte Referral Hospital, Western Oromia, Ethiopia. MATERIALS AND METHODS:Hospital-based cross-sectional study was employed from September 1, 2017, to October 30, 2017. Whole blood was tested for CrAg using Cryptococcal lateral flow assay (Immuno-Mycologics, Norman, OK, USA) according to the manufacturer's instructions. The collected data were analyzed using SPSS version 20 software. Binary logistic regression models were applied to assess the association between predictors and outcome variables at 95% CI. RESULT:Among the study participants, 115(62.8%) were females and the median age of the participants was 35 (IQR: 14) years. Majority, 169(92.3%), have been living with HIV for ? 5.6 years and 124 (67.8%) stayed on 2nd line ART for an average of 2.5 years. The overall prevalence of Cryptococcal antigenemia in the study participants was 7.7% (14/183). Being male [AOR, 95% CI: 4.78(1.14, 20.1)], poor adherence to ART [AOR, 95% CI: 0.12(0.03, 0.4)], occupational exposures to contaminated soil [AOR, 95% CI: 6.81(1.38, 33.4)], having non-separated house from chickens [AOR, 95% CI: 0.06(0.01, 0.51)], CD4 T+ cell levels ? 100 cell/?L [AOR, 95% CI: 6.57(1.9, 23.3)] and viral load >1000 copies/mL [AOR, 95% CI: 11.7(2.4, 57.8)] were significant predictors of Cryptococcal antigenemia (P? 0.05). CONCLUSION:The prevalence of Cryptococcal Antigenemia was significantly high in this study. Being male, occupational exposure to contaminated soil with avian droppings, CD4+ T cell levels <100 cell/?L and viral load >1000 copies/mL were significant predictors of Cryptococcal antigenemia. Therefore, public health measures, adherence to ART and early treatment are recommended.