ABSTRACT: Background: The world is facing the Coronavirus pandemic, which is highly infectious. Several measures have been put in place to prevent its spread among the population. However, for these preventive measures to be effective, the population requires appropriate and sufficient knowledge, attitude, and practices. Thus, a survey to assess knowledge, attitude, and self-reported practice toward measures for prevention of the spread of COVID-19 was conducted among Ugandans. Methods: This was a cross-sectional study conducted among during the lockdown in Uganda. An online structured questionnaire was used, applying a snowballing sampling approach for recruitment of participants 18 years and above and residing in Uganda. Data collection was done from 6th to 15th April 2020, during which 1,763 people participated. We analyzed all data using STATA 14.2, applying appropriate statistical tests. Results: Out of 1,763 participants, 80% were highly knowledgeable. For attitude, 72.4% reported following recommendations given by the Ministry of health to prevent the spread of COVID-19; 89.0% were worried about contracting COVID-19 and 73.3% agreed that COVID-19 can be cured and 99.3% reported good practice toward measures to prevent the spread of COVID-19. According to ordered logistic regression, health workers were 6 times more knowledgeable [aOR:6 (3.51–10.09), p < 0.001] followed by teachers [aOR:5.2 (2.6–10.32), p < 0.001]; students [aOR:3.2 (1.96–5.33), p < 0.001]. On the contrary, the drivers, business entrepreneurs, and security personnel had less knowledge. Conclusion: The results show that the participating Ugandans were knowledgeable and had a positive attitude and good practices. However, there is still a gap in knowledge among drivers, business entrepreneurs, and security personnel. Therefore, there is a need to mobilize the country's population to have the same degree of knowledge, which will have an impact on the attitude and practices toward prevention of the spread of COVID-19.