Several characteristic ions were observed during the direct analysis of a variety of both gram-negative and gram-positive intact bacterial cells by the matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) technique. The entire process, involving absolutely no sample processing, could be completed in less than ten minutes. A number of specific biomarkers, generated reproducibly for each type of cell from the corresponding mass spectrum, permitted the identification, as well as the distinction, of pathogenic bacteria from their non-pathogenic counterparts. In addition, individual strains of a specific organism could also be differentiated easily. Some of these biomarkers correspond to those observed earlier during the MALDI-MS analysis of protein extracts of the same bacteria. This approach, which can yield valuable data for rapid classification and detection of microorganisms, represents a substantial breakthrough for rapid screening of environmental as well as biological samples.