[The application of time-of-flight mass spectrometry with matrix activated laser desorption-ionization (MALDI-ToF) for identifying agents of glanders and melioidosis].

The article presents the results of application of developed methodological approach to identifying Burkholderia pseudomallei and Burkholderia mallei using direct mass spectrometry profiling of cellular proteins. The protocol of sampling preparation of cultures of melioidosis and glanders was optimized with taking in account characteristics of observation of requirements of biological safety for operations with pathogenic biological agents of pathogenicity group II. The dependence of quality of mass spectrums (number of individual peaks and their intensity) from medium of fermentation of microorganisms was evaluated. The characteristic mass spectrums of collection strains B.pseudomallei (5) and B.mallei (5) were obtained. The set of reference mass-spectrums was generated for identification data base S.A.R.A.M.I.S.TM (Anagnostec Gmbh.). The mentioned data base was used for identification of 43 strains of pathogenic Burkholderia. The opportunity of reliable identification of taxonomic belonging of examined microorganisms up to species’ level. The cluster analysis of obtained mass-spectrums of common cellular proteins of collection strains of pathogenic Burkholderia demonstrated grouping of examined strains according to their species’ belonging. The supplemented data base of mass-spectral characteristics hereinafter will permit applying express-identification of isolates suspicious for belonging to agents of melioidosis and glanders. The updated data base will become a basis for developing schemes of hemotyping of strains of Burkholderia using mass spectrometry technique.