Abstract Background: Leprosy is a chronic infectious disease of public health importance and one of the leading causes of permanent physical disability. The integration of leprosy service to the general health system in Ethiopia made leprosy patients to be seen by non-leprosy specialists which could lead to misdiagnosis and delay in treatment. In addition to the multi-drug treatment, patient self - care practice is crucial for the successful treatment of the disease. This study was aimed to assess the knowledge and self-care practices of leprosy patients and associated factors at ALERT leprosy referral Hospital in Ethiopia. Method: A cross-sectional study was conducted ALERT leprosy referral Hospital, Addis Ababa, Ethiopia. A total of 424 leprosy patients were interviewed using pre-tested structured questionnaires. The questionnaires included core points such as socio-demographic characteristics, knowledge of leprosy and self- care practices. Bloom's cut off point was used to describe the knowledge and self-care practice of the respondents and statistical significance was assessed at 95% confidence interval with 5% of level of significance. Result: The knowledge score of the respondents was poor for 276 (65.1%) and good for 148 (34.9%). The level of knowledge was significantly varied within age groups (P= 000), sex (P=000), marital status (p=0.003), educational status (p=000) and income (p-000). More than three-fourth (77.4%) of interviewed patients had poor self-care practice and only less than a quarter (22.6%) of patients had good self-care practice score (P=000). Age (p=0.002), Previous disability due to leprosy (P=000), Knowledge of leprosy (p=0.038) and income (P=0.028) significantly associated with poor self-care practice. On the other hand, educational status, sex, marital status and health education did not associated with leprosy self-care practice. Conclusion: Although, leprosy treatment, disability prevention and rehabilitation program run in the country under general public health care service for decades, poor leprosy self-care practice and poor leprosy knowledge had been confirmed in this study. Therefore, the leprosy service program should re-visit its strategy and mode of delivery to improve the leprosy knowledge and leprosy self-care practices of patients.