Healthcare workers (HCWs) exposed to Coronavirus disease 2019 (COVID-19) are not immune to stressors. This study aimed to explore the prevalence of posttraumatic stress symptoms (PTSS) among HCWs during the COVID-19 epidemic and investigate the associations among negative coping, fatigue and PTSS. A total of 507 HCWs from Anhui province enrolled in the study and completed the cross-sectional survey including demographic data, Simplified Coping Style Questionnaire (SCSQ), 14-item Fatigue Scale (FS-14), and PTSD Checklist-civilian Version (PCL-C). Univariate linear regression, Pearson correlation and Mackinnon's four-step procedure were performed in the statistical analysis. Results indicated that the prevalence of PTSS among HCWs during the pandemic was 24%. Univariate linear regression showed HCWs aged 31-40 years exhibited significantly higher scores of PTSS than those aged 51-60 (beta = 0.20, 95% CI: 0.59 to 9.41). Having at least one child was associated with a higher risk of developing PTSS (beta = 0.01, 95% CI: 0.36 to 5.45). Negative coping and fatigue were positively correlated with all three PTSS (all P < 0.001), including re-experiencing, avoidance and hyper-arousal. Fatigue has mediated the association between negative coping and PTSS among HCWs during the pandemic (ab = 0.09, SE = 0.03, bootstrap 95% CI: 0.04 to 0.14). A considerable proportion of HCWs was traumatized during the COVID-19 outbreak. Hence, the institutions should screen out and pay close attention to HCWs who tend to use negative coping (e.g., withdrawal thinking, distraction and blaming others) and arrange work scientifically to avoid overfatigue and PTSS amid the public health crisis.