Burnout among healthcare workers is a global concern with significant implications for both the well-being of the workforce and the quality of patient care. This facility-based cross-sectional study aimed to identify factors associated with burnout among healthcare workers in central Uganda. The study, conducted between June and July 2023, utilized a cross-sectional design involving physicians, nurses, and technicians. The study covered both public and private hospitals in central Uganda, incorporating urban and rural settings. A sample size of 550 healthcare workers was selected using a simple random sampling. Data collection involved a socio-demographic survey, the Professional Quality of Life (ProQOL-5). Descriptive statistics, Pearson Chi-square test, and ordinal regression models were employed to analyze demographic factors associated with burnout. Of 548 participants, 218(39.8%) experienced high burnout levels. Factors significantly associated with high burnout levels included having over 10 years of work experience (OR: 2.04, 95% CI: 1.12-3.73), working more than 40 hours per week (AOR: 4.46, 95% CI: 1.20-16.62), lack of management support (AOR: 14.45, 95% CI: 3.83-54.56), not experiencing workplace violence (AOR: 2.22, 95% CI: 1.31-3.76), and reporting inadequate sleep (AOR: 6.96, 95% CI: 3.86-12.57). Addressing burnout among healthcare workers in central Uganda requires targeted interventions tailored to the specific challenges faced in the region, including workload distribution, managerial support enhancement, violence prevention strategies, and promotion of adequate sleep. Urgent attention to these factors is essential for enhancing the well-being of healthcare professionals and maintaining quality patient care.