| Summary: | Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-TOF MS) technology is currently increasingly used in diagnostic laboratories as a cost effective, rapid and reliable routine technique for the identification and typing of microorganisms. In this study, we used MALDI-TOF MS to analyze a collection of 160 strains belonging to the <i>Bacillus cereus</i> group (57 <i>B. anthracis</i>, 49 <i>B. cereus</i>, 1 <i>B. mycoides</i>, 18 <i>B. wiedmannii</i>, 27 <i>B. thuringiensis</i>, 7 <i>B. toyonensis</i> and 1 <i>B. weihenstephanensis</i>) and to detect specific biomarkers which would allow an unequivocal identification. The Main Spectra Profiles (MSPs) were added to an in-house reference library, expanding the current commercial library which does not include <i>B. toyonensis</i> and <i>B. wiedmannii</i> mass spectra. The obtained mass spectra were statistically compared by Principal Component Analysis (PCA) that revealed seven different clusters. Moreover, for the identification purpose, were generated dedicate algorithms for a rapid and automatic detection of characteristic ion peaks after the mass spectra acquisition. The presence of specific biomarkers can be used to differentiate strains within the <i>B. cereus</i> group and to make a reliable identification of <i>Bacillus anthracis</i>, etiologic agent of anthrax, which is the most pathogenic and feared bacterium of the group. This could offer a critical time advantage for the diagnosis and for the clinical management of human anthrax even in case of bioterror attacks.
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