Background:Routine eye examination plays a vital role in detecting diabetic retinopathy in its earliest stage before the onset of blindness. Patients’ knowledge about the nature and the consequences of diabetic retinopathy and routine eye checkup helps in timely identification and early treatment. However, there is limited evidence on knowledge of patients with diabetes mellitus on diabetic retinopathy and their eye check-up practices in Ethiopia.The aim of this study was to assess knowledge, eye check-up practice and associated factors of diabetic retinopathy among adult diabetic patients at Debark hospital, Northwest Ethiopia.Methods: Institution based cross – sectional study was conducted at Debark hospital, Northwest Ethiopia, from April 20/2018- May 20/2018. A pretested interviewer administered structured questionnaire was used to collect data among 230 diabetic patients aged 18 years and above. Data were entered in to Epi Info version 7 and exported to SPSS version 20 for analysis. Bivariable and multivariable binary logistic regression analyses were done. Odds ratio with 95% confidence level was determined and variables with p–value of < 0.05 were considered as statistically significant.Result: Out of 238 sample 230 were participated, among this, 119 (51.7%) were males. The mean age of the respondents was 49 (SD ±17.6) years. One hundred nine (47.4%) participants had good knowledge and 91 (39.6%) had good eye check-up practice. Urban residence [AOR= 2.65;95% CI: 1.16-6.07)]), monthly income of 3501-8000 birr [AOR=4.54;(1.31-15.7)], type II diabetes mellitus [AOR=3.9;(1.6-9.6)], duration of diabetes (6-12 years[AOR=4.4;(1.4-13.5)]), history of eye disease [AOR=5.5;(2.3-13.0)] were associated with good knowledge. Similarly, longer duration of diabetes (13-25 years [AOR=3.77; (1.05-13.5)]) and history of eye disease [AOR=2.47; (1.09-5.62)] were associated with good eye check-up practice. Conclusion: The proportion of good knowledge about diabetic retinopathy among diabetic patients at Debark hospital was fair (47.4%) and good eye check-up practice (eye examination at least once in the past year) was low (39.6%). Longer duration of diabetes and history of eye disease were identified as positive factors for good knowledge and eye check-up practice. Knowledge and regular eye check-up practice needs to be enhanced through provision of appropriate health education.