Aim: During the 2019 coronavirus disease (COVID-19) pandemic, healthcare workers experience intense levels of workload and pressure. The exact psychological and mental impacts of this pandemic on healthcare workers and relevant factors remain to be elucidated. Therefore, this study aimed to investigate the stress and anxiety of healthcare workers due to COVID-19. Material and Methods: This prospective multi-centered survey-based study was conducted on 205 participants working in the emergency room (ER) of two hospitals between May 15 and May 22, 2020. Besides demographic data such as age, gender, working history in ER, marital status and presence of children, the Perceived Stress Scale (PSS) and the Beck Anxiety Inventory (BAI) were used in the questionnaire. Data were analyzed by either the Kruskal-Wallis test and Mann-Whitney U test, Chi-square test or Fisher's exact test. Results: Most of the participants were nurses (35.5%), and 53.2% of all participants were male. The rate of COVID-19 infection among physicians and nurses was significantly higher than among personnel in other occupations. The rate of the willingness of physicians and nurses to receive psychiatric support was significantly higher than that of other personnel (p < 0.05). Physicians and nurses had higher BAI and PSS scores than other staff, and scores of nurses were significantly higher than physicians (p < 0.05). Discussion: During the pandemic, the adverse psychological and mental impacts of COVID-19 were prevalent among the healthcare workers. Adequate measures at both organizational and national levels are required to improve the psychological state of frontline healthcare workers during the COVID-19 pandemic.