Introduction: Depression is a serious issue affecting healthcare workers and is a leading cause of disability for both genders. Furthermore, it is one of the leading causes of mortality and morbidity, responsible for 4.4 percent of global disability. An estimated 350 million people are currently living with depression worldwide. Objectives: to estimate the prevalence of depression among healthcare workers in Khartoum State in 2022 and determine the associated factors. Methods: A cross-sectional survey was conducted among healthcare workers in Khartoum State, Sudan, in 2022 using a self-administered electronic questionnaire. Depression was screened using the self-reporting questionnaire (PHQ-9). Descriptive statistics in the form of frequencies and percentages were used to display the data. Odds ratios (ORs) with a 95% confidence interval were estimated using bivariate and multivariate logistic regression analysis to determine associations between depression and related factors. Results: A total of 341 valid responses were received, with a mean age of 33.91. The overall prevalence of depression (PHQ-9 > 8) was 258 (75.6%). The prevalence was significantly associated with marital status (single and divorced), occupation (psychologist), and working department (Emergency Department), showing a p-value of <0.041. Conclusion: Depression is a serious mental health disorder that affects all people, including healthcare workers, and is a growing problem in Sudan. To address this, healthcare organizations must implement policies and strategies to reduce inequality and protect healthcare workers. A multidisciplinary approach that includes mental health professionals, the Ministry of Health, and universities is needed to prioritize mental health issues and ensure quality care and the overall well-being of both healthcare workers and patients.