Background: This study aims to assess COVID-19 related knowledge and practice among the Egyptians in Upper Egypt and to identify barriers that hinder adherence to these preventive measures. Design and methods: We conducted a cross-sectional study using a structured questionnaire. Data was collected from 21 to 30 August 2020, via both online survey and personal interviews using a non-probability self-nominated sample. Results: A total of 731 participants completed the questionnaire. The mean age was 32.1± 2.1 and 64.3% were females. The main sources of knowledge were, Internet and Social Media followed by TV then family and friends. More than 96% of the respondents knew the origin, nature, the main symptoms, and the modes of transmission of COVID-19, however, 37.6% of them thought that COVID-19 patients must develop symptoms. 75.8% and 73.6% of the participants respectively covered their nose and mouth during sneezing and washed their hands regularly, 65.4% wore masks in crowded places, while only 31.1% and 30% of them respectively avoided touching their faces or shaking hands with friends. Knowledge and practice were positively correlated and both were linked to younger age and higher education and the female gender was also a predictor of better practice. The most common perceived barriers to adherence to preventive measures were feeling uncomfortable, forgetfulness then financial causes. Conclusions: the study subjects were aware of COVID-19 and its preventive measures however adherence to some of these measures was not prevalent amongst them, indicating an urgent need of addressing and targeting barriers that hinder adherence to COVID-19 preventive measures in the future policies to prevent the spread of COVID-19 in Egypt.