Differentiation of Streptococcus pneumoniae from nonpneumococcal streptococci of the Streptococcus mitis group by matrix-assisted laser desorption ionization-time of flight mass spectrometry

Abstract

The differentiation of species within the Streptococcus mitis group has posed a problem in the routine diagnostic microbiology laboratory for some time. It also constitutes a major weakness of recently introduced matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) fingerprinting systems. As the phylogenetic resolution of the spectral similarity measures employed by these systems is insufficient to reliably distinguish between the most closely related members of the group, the major pathogen Streptococcus pneumoniae is frequently misidentified. In this study, a comparative analysis of MALDI-TOF spectra of several species from the S. mitis group has been performed in order to identify single peaks that could be used to improve mass spectrometry-based identification of the respective species. A characteristic peak profile could be identified that unambiguously distinguished the 14 S. pneumoniae isolates studied from 33 nonpneumococcal isolates of the S. mitis group. In addition, specific peak combinations could be assigned to other members of the group. The findings of this study suggest that it is possible to distinguish different species of the S. mitis group by close analysis of their mass peak profiles.

Fig 1

Individual m/z values as they are visible on the flexAnalysis software. Spectra are overlaid to ensure consistency with measured intensities. The chosen spectra were from strains with sequence-confirmed identification. S. pneumoniae, pink; S. pseudopneumoniae, green; S. oralis, black; S. sanguinis, blue; and S. mitis, red. (A) m/z 2,625; (B) m/z 2,911and 2,937.5; (C) m/z 5,253; (D) m/z 5,824; (E) m/z 5,877; and (F) m/z 6,955.