Background: The coronavirus disease 2019 (COVID-19) pandemic has negatively impacted public health systems worldwide and created anxiety and stress among communities, resulting in the stigmatization of patients infected with the virus. Stigmatization of individuals who are sick or thought to be infected has a long history and can lead to discrimination and prejudice. This study aims to evaluate the prevalence of COVID-19-related stigma in Jordan, assess the relationship between stigma and the quality of life (QoL) in healthcare workers, and identify possible measures to decrease stressful events. Understanding the psychological effects of healthcare workers' jobs and reducing their burden is essential to improving medical outcomes and the QoL of patients.Methodology: This cross-sectional study was conducted in three primary hospitals in Amman, Jordan, from July to December 2021. Healthcare workers were recruited through convenience sampling and completed a self-administered questionnaire, which included demographic information, a validated COVID-19 stigma questionnaire, work conditions during the pandemic, the Depression, Anxiety and Stress Scale - 21 Items (DASS-21) questionnaire to measure depression, anxiety, and stress, and the EUROHIS-QOL questionnaire to assess the QoL. Data were analyzed using descriptive and inferential statistics, including chi-square tests and post hoc analysis. The study was approved by the institutional review board, and participation was voluntary and confidential.Results: The study was conducted among 683 healthcare workers in Jordan, with 77.7% based in the capital city, Amman. Most of the participants were between 18 and 30 years of age, and slightly more than half were female. The study found that 38.1% of healthcare workers would not take the COVID-19 vaccine once it became available. More than half (56%) reported experiencing stress, 61% reported anxiety, and 65% reported depression during the pandemic. Internal medicine specialties and frontline nurses reported the highest levels of stress, and healthcare workers with greater exposure to COVID-19 patients reported higher levels of anxiety and stress. Only 3% of participants reported experiencing stigmatization (p=0.043), with low-income participants reporting it more frequently. Stigmatization was significantly correlated with feelings of depression, anxiety, and stress (p<0.001).Conclusion: Healthcare workers’ mental well-being has been affected negatively by the COVID-19 pandemic, resulting in depression, anxiety, and stress. Widespread mental surveillance for healthcare workers is crucial to protect healthcare workers from psychological issues and to improve the healthcare service provided to patients. Stigma among healthcare workers can be a major factor that may increase depression, anxiety, and stress.