Abstract: Objective To investigate the prevalence and clinical characteristics of epstein-barr (EB) virus infection in the hospital and to provide scientific theoretical basis for the prevention and treatment of EBV infection and related diseases. Methods A retrospective study was conducted among EB infected patients in our hospital from October 2015 to September 2016. The relationship between detection results and clinical data of patients were analyzed. Results The plasma samples from 1 313 patients were collected and detected for EB infection. 314 cases were found EBV-DNA positive with a total positive rate of 23.91%. The positive rates of female patients (21.60%) and male patients (25.27%) showed no significant difference ((口恶)²=2.26, P=0.133). The positive rate of EBV-DNA varied in different age groups, peaking at 1-10 and 61-70 years old. The positive rate altered by different seasons was highest (36.44%) in autumn and lowest in winter (13.00%). There was significant difference in the EB infection rate among different departments of the hospital. The top five departments with high infection rate of EB were pediatrics (40.91%), radiotherapy (39.30%), hematology (18.50%), infectious disease (16.98%) and intensive care unit (15.79%). The positive rate of EBV-DNA in different diseases was different. Conclusions Infection rate of EB virus in our hospital is comparable with that in other regions of China. The infection rate varies in different seasons, ages and departments in the hospital. The clinical manifestations of EB virus infection are complex and varied.