Abstract: Objective The aim of this study was to screen serum biomarkers in patients with acute leukemia by using surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technique. Methods Proteomic spectra were generated by mass spectrometry in 158, including 44 acute lymphocyte leukemia patients and 36 acute myeloid leukemia, 78 healthy control. 158 spectra obtained were used to train and develop a decision tree classification algorithm. Results A total of 7 distinguished proteomic peaks were detected in acute lymphocyte leukemia and were used to build a proteomic pattern. The results yielded a sensitivity of 88.6％（39/44）、specificity of 82.9％（29/35）；A total of 8 distinguished proteomic peaks were detected in acute myeloid leukemia，7 peaks were used to build a proteomic pattern. The results yielded a sensitivity of 86.1％（31/36）、specificity of 95.3％（41/43）. Conclusion SELDI-TOF-MS offers a unique platform for the proteomic detection of acute leukemia. It also offers a noninvasive method to further study the proteomic changs in the development and progression of leukemia.