BackgroundCoronavirus disease 2019 (COVID-19) has been a global public health crisis since December 2019. Health care professionals (HCP) are highly vulnerable for contracting the disease. Due to the absence of known treatment, prevention is the best option for controlling its transmission.ObjectiveThe study aimed to assess COVID-19 prevention practices and determinant factors among HCP working in hospitals of South Gondar ZoneH, Northwestern Ethiopia.MethodsInstitution-based cross-sectional study design was used from July 1 to 30, 2020 among 372 HCPs. The participants were selected on random sampling technique basis. Data were entered into Epi data version 3.1 and exported to Statistical Package for Social Science (SPSS V.25) software for data cleaning and analysis. Binary logistic regression analysis was used to assess the association between independent variables and prevention practice of COVID-19.ResultsA total of 372 HCPs participated in the study with a response rate of 94.9%. Less than one-third 112 (30.1%) of the participants were females. About 308 (82.8%), 285 (76.6%), and 224 (60.2%) of the participants had good knowledge, positive attitude, and good prevention practice towards COVID-19, respectively. Being male (AOR =2.68; 95% CI (confidence interval): 1.50,4.81), 2–5 years working experience (AOR= 4.61; 95% CI: 1.70,12.47), greater than five years working experience (AOR= 5.86; 95% CI: 2.01,17.05), age of 31–40 years old (AOR= 2.49; 95% CI; 1.41,4.41), above 40 years (AOR= 6.94; 95% CI: 2.33–20.71), use of COVID-19 guideline (AOR= 4.79; 95% CI: 2.17–10.53), and using peers as sources of information about COVID-19 (AOR= 2.06; 95% CI: 1.07–3.94) were factors of COVID-19 prevention practices.ConclusionLess than two-thirds of the participants had good COVID-19 prevention practices. Sex, work experience, age, use of COVID-19 guideline, and using peers as sources of information were factors of COVID-19 prevention practices. Hence, continuous information dissemination and experience sharing on COVID-19 prevention should be done continuously.