Abstract: Objective　To explore the clinical characteristics and relevant factors affecting treatment and prognosis of TCF3-PBX1 positive acute lymphoblastic leukemia (ALL). Methods　The clinical data of 29 children with newly diagnosed TCF3-PBX1 positive ALL from August 2006 to August 2015 were analyzed retrospectively. The expression level of TCF3PXB1 fusion gene was monitored by regular quantitative reverse transcription polymerase chain reaction. The factors influencing prognosis in children with TCF3-PBX1 positive ALL were analyzed. Results　There were 29 children (16 males and 13 females) with a median age of 8 years (9 months to 16 years). The most common immunophenotype was pre-B cell type (pre-B) (58.6%). The karyotype analysis showed that unbalanced translocation was more common (41.4%). The complete remission rate was 100% on thirty-third day in 29 children, but the minimal residual disease (MRD) was not completely negative. Three cases were relapsed, all of whom were MRD positive. Cox multivariate regression analysis showed that age was an independent risk factor for 5 year overall survival (P<0.05). The 5 year overall survival rate and disease-free survival rate were (82±8)% and (81±7)% respectively. Conclusions　Childhood TCF3-PBX1 positive ALL is a highly heterogeneous disease with high rate of complete remission and good long-term efficacy. The risk stratification and individualized treatment is the key to improve the cure rate.