Abstract：Objective To investigate the incidence and related risk factors of Epstein-Barr virus （EBV） infection after pediatric hematopoietic stem cell transplantation, and to provide evidence for the prevention and treatment of pediatric hematopoietic stem cell transplantation. Methods A retrospective study of 48 pediatric patients undergoing hematopoietic stem cell transplantation for malignant and non-malignant disease at our hospital from January 2014 and December 2020. All of them received pulp removal preconditioning and reduced-intensity preconditioning. Results  （1） The positive rate of EBV infection was 12.50% （6/48） by quantitative PCR before transplantation. The positive rate of EBV infection was 41.67% （20/48）. Within 3 months after transplantation, there was 1 case of lymphocyte proliferative diseases （2.08%）. （2）The positive rate of EB infection rate was 77.08% （37/48）. The positive rate of EBV infection in the negative children who were not infected before transplantation was 50.00% （24/48） after transplantation, although the infection rate was lower than that of the former, there was no significant difference between them （P＝0.058）. Conclusion The guidelines for monitoring EB virus PCR in children after hematopoietic stem cell transplantation are not yet perfected, and routine viral PCR monitoring results can improve healthcare utilization and iatrogenic blood loss, and effectively prevent EBV infection in children after hematopoietic stem cell transplantation.