Background: Stroke is ―rapidly developing clinical signs of focal or global disturbance of cerebral functions, lasting more than 24 hours or leading to death, with no apparent cause other than that of vascular origin.‖ The prevalence of stroke is decreasing in the developed countries while it is increasing in low level and middle income countries. Stroke Deaths in Ethiopia reached 39,571 or 6.23% of total deaths. Objective: The study was aimed to assess determinants of stroke among patients in referral hospitals of Amhara regional state, Ethiopia 2020 Method: Institutional based case control study was conducted among 132 stroke and 264 non-stroke patients who attended the referral hospitals of Amhara regional state from March 1/ 2020 to April 15/2020. The hospital and respondents were selected by simple random sampling and systematic random sampling respectively. The data were collected by face to face interview using Amharic version questionnaire. Data analysis was done by Statistical package for social science version 23. Descriptive statistics were done using frequency distribution, percentage, table and graphs to describe sociodemographic, behavioral and clinical factors. Bivariable and multi-variable binary logistic regressions were used to check explanatory variables having association with the dependent variables. Backward conditional logistic regression was used to identify factors associated with stroke. The model fitness test was evaluated using Hosmer and lemeshow goodness-of-fit. Results: The study included 419 respondents with 95% response rate. The study revealed that, previous history of alcohol (AOR=1.61, 95% C.I: 1.15-2.78), Self-reported stress (AOR=1.82, 95% C.I: 1.15- 2.88), unrestricted dietary salt intake (AOR= 1.89, 95% C.I: 1.19-2.99), Migraine disease (AOR=1.72, 95% C.I: 1.04-2.83) and cardiac disease (AOR= 1.85, 95% C.I: 1.14-2.99) were determinants of stroke. On the other hand regular physical exercise (AOR=0.496, 95% C.I: 0.29-0.84) was preventive factor of stroke. Conclusion and Recommendation: The finding from this study highlights the associations of stroke with previous history of alcohol, Self-reported stress, unrestricted dietary salt intake, Migraine disease and cardiac disease. So implementing preventive strategies on those risk factors is needed to reduce the risk of stroke. Key words: - Stroke, determinants, Ethiopia