Abstract: 　Objective　To investigate the distribution of pathogenic microbes in blood culture and analysis of drug resistance of common pathogens in children with acute leukemia. Methods　BACTE-FX system made by BD company was applied for the blood culture of children with acute leukemia. API or VITEK-2 Compact bacterial identification system was used for identification of bacteria. Antimicrobial susceptibility testing was performed by disk diffusion method. Results　Among 6400 blood culture samples in children with acute leukemia, 713 cases were positive at the rate of 11.1%. Among 655 strains first isolated from the patient, Gram-positive bacteria were 52.5% (344/655) and dominated by Staphylococcus. Strains resistant to vancomycin and linezolid were not found. 278 strains (42.5%) were Gram-negative bacteria , and S.marcescens and E.coli were dominated. Gram-negetive bacteria showed high antibacterial activity to imipenem with the sensitive rate of 100%. 33 strains (5%) were fungi. Conclusions　Staphylococcus, S.marcescens, and E.coli were the most important bacteria in blood culture of children with acute leukemia. We can offer effective help for clinical treatment with the susceptibility testing.