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This study aimed to assess the level of knowledge, attitude, and practice regarding the coronavirus disease among Iranian rural and urban healthcare workers. The study population consisted of all healthcare experts and staff working in 38 rural health centers in Iran. An online questionnaire was distributed in November 2021. The anonymous questionnaire was comprised of a series of questions about demographic information; knowledge about COVID-19, the attitudes and preventive practices toward COVID-19. The majority of the participants (92.6%) had appropriate knowledge about COVID-19 disease. Most of the participants (77.2%) had a good attitude toward COVID-19 and 60.3% showed good practice. There was a significant, strong, and positive correlation between knowledge and attitude scores (p value