Objective To study immunophenotype and clinical characteristics of acute myeloid Leukemia (AML) with FLT3-ITD positive. Methods The immunophenotype and laboratory characteristics of FLT3-ITD+and FLT3-ITD-acute myeloid leukemia patients admitted to our hospital from June 2015 to June 2017 were retrospectively analyzed. The clinical outcomes and survival of the two groups after chemotherapy were compared. Results The expression of CD56, CD33, CD11b, CD7 in FLT3-ITD+AML patients was much higher than that in FLT3-ITDAML patients, while the expressions of CD34 and CD117 were much lower (P < 0.05); the white blood cell count and bone marrow leukemia cells in FLT3-ITD+AML patients increased much more than those in FLT3-ITD-AML patients (P < 0.05), while there was no statistical difference in hemoglobin and platelet count (P > 0.05); the rate of treatment objective response (OR) in FLT3-ITD+AML patients was much higher than that in FLT3-ITD-AML patients (P < 0.05); Patient-free survival and overall survival in FLT3-ITD-AML patients were much higher than those in FLT3-ITD+AML patients (P < 0.05). The expressions of CD56, CD34, CD11b, CD7, CD34 and CD117 in FLT3-ITD+patients with complete remission were significantly lower than those in patients without remission (P < 0.05). Conclusions The abnormal expression of leukemia cell antigen, high peripheral blood leukocytes and bone marrow primordial cells, low complete remission rate and poor prognosis in patients with FLT3-ITD+AML are the poor prognostic factors of AML. Early detection of FLT3-ITD mutation and immunotyping in AML patients has important clinical significance in guiding treatment and judging prognosis.