We examined the gene expression profiles of two independent cohorts of patients with acute myeloid leukemia [n= 247 and n= 214 (younger than or equal to 60 years)] to study the applicability of gene expression profiling as a single assay in prediction of acute myeloid leukemia-specific molecular subtypes. The favorable cytogenetic acute myeloid leukemia subtypes, ie, acute myeloid leukemia with t (8; 21), t (15; 17) or inv (16), were predicted with maximum accuracy (positive and negative predictive value: 100%). Mutations in NPM1 and CEBPA were predicted less accurately (positive predictive value: 66% and 100%, and negative predictive value: 99% and 97% respectively). Various other characteristic molecular acute myeloid leukemia subtypes, ie, mutant FLT3 and RAS, abnormalities involving 11q23,− 5/5q-,− 7/7q-, abnormalities involving 3q (abn3q) and t (9; 22), could not be correctly predicted using gene expression profiling. In conclusion, gene expression profiling allows accurate prediction of certain acute myeloid leukemia subtypes, eg those characterized by expression of chimeric transcription factors. However, detection of mutations affecting signaling molecules and numerical abnormalities still requires alternative molecular methods.