Background: The pathological report suggested severe immune injury in corona virus disease 2019 (COVID-19) patients. Epstein-Barr virus (EBV) reactivation has been reported in immunocompromised individuals. We aimed to detect EBV coinfection in COVID-19 patients. Methods: In this single-centered, retrospective, observational study, COVID-19 patients were enrolled in our study. The characteristics of the demographic, symptoms, signs, laboratory and CT results and clinical outcome were collected. Data were compared between EBV seropositive and seronegative COVID-19 patients. Findings: Of 196 patients, 62 COVID-19 patients were included in our study. The median age was 37 years, with 32 (51 · 6%) females. Among these COVID-19 patients, 35 (56 · 5%) patients were seropositive for EBV viral capsid antigen (VCA) IgM antibody. EBV seropositive COVID-19 patients had a 3 · 64-fold risk of having a fever symptom than EBV seronegative (95%CI, 1 · 26-10 · 51; P=0 · 02). C-reactive protein (CRP) in EBV seropositive COVID-19 patients were higher than EBV seronegative patients (P=0 · 01). The proportion of CD8 (CD8%) was statistically significant lower in EBV seropositive COVID-19 patients (P=0 · 048). The CD4/CD8 in EBV seropositive COVID-19 patients was lower than that in EBV seronegative patients (P=0 · 046). 25 (40 · 3%) were discharged before February 29, 2020. The median recovery time in EBV seropositive COVID-19 patients (24 days) was higher than that in EBV seronegative COVID-19 patients (19 days), while the difference was not significant (P=0 · 07). Interpretation: EBV acute infection was found in COVID-19 patients. EBV seropositivity was associated with fever, increased inflammation and cellular immune response in COVID-19 patients. EBV reactivation should be considered in treating COVID-19. Funding Statement: None. Declaration of Interests: The authors confirmed that no competing financial interests exist. Ethics Approval Statement: The study was approved by the ethical committee board of Renmin Hospital of Wuhan University (WDRY2020-K073).