|Fast and accurate species identification of microorganisms from positive blood cultures is of high importance to guide the subsequent antibiotic therapy. Currently, the identification takes up to two days because of the necessity for sub-cultivation and biochemical identification. We present a simple protocol which can shorten identification time to minutes. Microorganisms could be isolated and prepared for MALDI-TOF MS measurement in less than 30 minutes. Tests in several clinical microbiology laboratories did reveal identification rates of 69.4 – 88.3% which could be further increased by adopted spectra processing and result interpretation.