The MALDI-TOF mass spectrometry is a new kind of generation of identification systems for microorganisms. Within a few minutes, bacterial or fungal proteins are being analysed by a mass spectrum through a database. To date routine methods used for identification of microorganisms are gram staining and biochemical methods. It takes 24 hours to get a result. This work is dealing with the basics of MALDI-TOF mass spectrometry and the comparison of the MALDI-TOF mass spectrometry instrument VITEK MS and the biochemical reactions based analyzer VITEK 2. For the measurements, routine representative grampositive bacteria are being used. The sample pool of my research includes Coagulase-negative staphylococci, Enterococcus faecium, Enterococcus faecalis, and Streptococcus agalactiae. Among 159 analysed samples, 153 matched completely. Five samples matched with conditions and one sample was being evaluated on the basis of a low probability value of VITEK 2, as not compliant. With these results, the rapid identification and the low identification costs, it can be concluded that the MALDI-TOF mass spectrometry will play an important role within the microbiological diagnosis.