PurposeThe stress of conscience occurs in stressful situations that are often encountered in healthcare, leading to an uncomfortable conscience in healthcare workers. They may not be able to control their emotions and feel burnout. Therefore, the aim of this study is to evaluate the effect of stress of conscience on burnout in healthcare workers.MethodsThis cross‐sectional study was performed with 602 healthcare workers between November 2020 and January 2021 in Turkey. Data were collected using a sociodemographic form, Stress of Conscience Scale, and Maslach Burnout Inventory.ResultsA positive and high‐level relationship was found between the stress of conscience and burnout (r = 0.603, p < 0.001). The stress of conscience accounted for 36% of burnout. Also, female healthcare workers have high stress of conscience and burnout scores.ConclusionThis study is important in terms of realising the stress and burnout of healthcare workers and for health managers to recognise the stress types of their workers. The increase in the level of burnout of healthcare workers is associated with their stress of conscience. Using the results of this study, national and international consensus can be obtained to reduce the impact of the stress of conscience on healthcare workers.