Abstract: Objective：To investigate the mutational features of FLT3-ITD , CEBPA, NPM1,DNMT3A, and KRAS in childhood patients with acute myeloid leukemia (AML) and their prognostic significance. Methods：A total of 217 newly diagnosed AML childhood patients were enrolled in the study. Their bone marrow samples were collected, and the mutations of FLT3-ITD, CEBPA, NPM1, DNMT3A, NRAS and KRAS were detected by PCR and Sanger sequencing. The relationships between these mutations and clinical data were evaluated. Results：The mutation frequencies of NRAS, CEBPA, FLT3-ITD, KRAS, NPM1 and DNMT3A were 11.9%, 10.0%, 5.7%, 3.0%, 1.4% and 0, respectively. The KRAS mutation existed only in FAB type M5 while the NPM1 mutation only in type M2. The peripheral WBC counts (×10⁹/L) in the patients with the FLT3-ITD mutation were significantly higher than that without the mutation ［104.0 (19.8, 201.0) vs 11.4 (3.8, 38.7), Z=-3.061, P=0.002］. The ages of the patients with KRAS mutation were significantly younger than that without the mutation ［2.0 (1.0, 3.3) year-old vs 7.0 (3.0, 10.0) year-old, Z=-2.28, P=0.005］. The overall survival (OS) rates in the patients with FLT3-ITD mutation were significantly shorter than that without the mutation (25.0% vs 52.5%, χ=4.993, P=0.026), and the eventfree survival (EFS) rates tended to reduce (33.3% vs 60.6%, χ=3.750, P=0.053). There was no significant difference in prognosis between the patients with RAS mutation and without mutation. Conclusion: FLT3-ITD mutation may be a biomarker for predicting the prognosis of pediatric AML.