title: Application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the study of Helicobacter pylori.
abstract: The characteristics of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry based investigation of extremely variable bacteria such as Helicobacter pylori were studied. H. pylori possesses a very high natural variability. Accurate tools for species identification and epidemiological characterization could help the scientific community to better understand the transmission pathways and virulence mechanisms of these bacteria. Seventeen clinical as well as two laboratory strains of H. pylori were analyzed by the MALDI Biotyper method for rapid species identification. Mass spectra collected were found containing 7-13 significant peaks per sample, and only six protein signals were identical for more than half of the strains. Four of them could be assigned to ribosomal proteins RL32, RL33, RL34, and RL36. The reproducible peak with m/z 6948 was identified as a histidine-rich metal-binding polypeptide by tandem mass spectrometry (MS/MS). In spite of the evident protein heterogeneity of H. pylori the mass spectra collected for a particular strain under several cultivations were highly reproducible. Moreover, all clinical strains were perfectly identified as H. pylori species through comparative analysis using the MALDI Biotyper software (Bruker Daltonics, Germany) by pattern matching against a database containing mass spectra from different microbial strains (n = 3287) including H. pylori 26695 and J99. The results of this study allow the conclusion that the MALDI-TOF direct bacterial profiling is suited for H. pylori identification and could be supported by mass spectra fragmentation of the observed polypeptide if necessary.
authors.affiliation: Research Institute of Physical-Chemical Medicine, Malaya Pirogovskaya st. 1a, Moscow 119992, Russia. ilinaen@gmail.com
authors.forename: Elena N
authors.initials: EN
authors.lastname: Ilina
authors.forename: Alexandra D
authors.initials: AD
authors.lastname: Borovskaya
authors.forename: Marina V
authors.initials: MV
authors.lastname: Serebryakova
authors.forename: Vera V
authors.initials: VV
authors.lastname: Chelysheva
authors.forename: Kuvat T
authors.initials: KT
authors.lastname: Momynaliev
authors.forename: Thomas
authors.initials: T
authors.lastname: Maier
authors.forename: Markus
authors.initials: M
authors.lastname: Kostrzewa
authors.forename: Vadim M
authors.initials: VM
authors.lastname: Govorun
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D016480:Helicobacter pylori / Q000737:chemistry* / Q000145:classification* / Q000302:isolation & purification; D006801:Humans; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*