Until now, bacterial and fungal identification has been based on biochemical characterization of microorganisms. The Matrix-Assisted Laser Desorption/Ionization Time of Flight Spectrometry (MALDI-TOF MS) has recently been developed in clinical microbiology laboratories. This new technology allows a rapid, accurate and less expensive identification of bacterial and fungal colonies grown on agar media. First, we have shown that the direct identification of bacteria grown in liquid media such as blood cultures was possible, without waiting for a subculture on solid media. Since the diagnosis is available on the first day, the presumptive antimicrobial treatment can be rapidly adapted according to the usual resistance phenotype of the microorganism. We have then searched to identify the biomarkers used for the identification of bacteria, using Neisseria meningitidis as a model. Comparing the genome and the proteome of sequenced strains allowed us to identify the ribosomal proteins as thoses involved in the MALDI-TOF MS diagnosis. Ribosomal proteins are very abundant and are very often used as epidemiological tools : their variations on the bacteria mass spectrum allows an intra-species differentiation of several strains. Finally we present encouraging preliminary results based on the detection of consistent ribosomal proteins directly visible on the mass spectrum that lead to the accurate identification of some very close species such as Streptococcus pneumoniae and Streptococcus mitis.