Evaluation of the Analytical Capabilities of MALDI-TOF Mass Spectrometry in the Molecular Typing of <i>Bacillus anthracis</i>

The aim of the work was to compare the discriminating capacity of the canSNP13 genotyping and MALDI‑TOF mass spectrometry methods based on the results of the study of anthrax pathogen strains belonging to the two main genetic lines A and B.Materials and methods. 73 Bacillus anthracis strains from the collection of microorganisms of the Stavropol Research Anti-Plague Institute of the Rospotrebnadzor were investigated. Protein profiling was carried out using a Microflex mass spectrometer, data processing – in the environment of the statistical programming language “R”.Results and discussion. It has been experimentally confirmed that the proposed approach for differentiating proteotypes of B. anthracis strains with a discrimination index of 0.952 exceeds the one for the canSNP typing method and is comparable to the discrimination index for the MLVA31 method. The correlation of the results of strain clustering during typing by MALDI-TOF mass spectrometry and canSNP genotyping reaches 95 % as regards the division into the main genetic lines A and B. The studied strains of anthrax microbe, which mostly fall under phylogenetic groups of lineage A, represent more than a dozen protein profiles, which may be due to differences in the level of protein expression in strains of each canSNP genotype. MALDI-TOF mass spectrometry makes it possible to obtain the results comparable with genetic tests, has a better discriminating capacity compared to canSNP typing, and is easier to perform

Development of Algorithm for Identification of Brucellosis Agent Cultures Using MALDI-TOF Mass-Spectrometry

Given are the results of direct protein profiling of brucella strain collection using matrix laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS). Standardized is the method for brucellosis agent culture disinfection and subsequent assessment. Obtained are 59 representative protein profiles of six major species ( B. melitensis, B. abortus, B. canis, B. ovis, B. neotomae ), which are included into electronic database of mass-spectra, integrated in Biotyper v 3.0 software (Bruker Daltonics, Germany). Analysis of mass-spectra of brucella strains under investigation revealed 17 genus-specific fragments within the mass range of 2000-20000 Da, the combination of which can be used for Brucella spp . identification. In addition, described are the fragments, specific for certain brucella species and strains, promising as markers for intraspecific differentiation. Based on the data received developed is a standardized algorithm of identification and differentiation of brucellosis agent cultures applying MALDI-TOF mass spectrometry

Analysis of Brucellosis Incidence and Molecular-Genetic Characteristics of Brucella Population in the Territory of the Russian Federation

An analysis of brucellosis incidence in Russia in 2013–2022 and the data on genetic diversity of Brucella melitensis and Brucella abortus populations isolated in Russia in 1939–2022 are provided in the review. Over the past decade, the epidemiological situation in Russia has been characterized as unstable against the background of persistent unfavorable conditions for brucellosis in cattle and small ruminants. During the period of 2013–2022 (9 months), 4298 epizootic foci as regards brucellosis in cattle (89164 sick animals) and 371 as regards brucellosis in small ruminants (13569) were registered. The largest number of epizootic brucellosis foci was recorded in the North-Caucasian and Southern Federal Districts. In 2013–2022, on average, 327 cases of brucellosis among people were registered annually, the incidence rate per 100 000 of the population was 0.24. Up to 70–90 % of brucellosis cases were detected in the south of the European part of the country. A trend towards deterioration of the situation on brucellosis in Volga (Penza and Samara Regions) and Central (Smolensk, Voronezh and Tula Regions) Federal Districts is observed. There is a connection between the intensity of epidemic manifestations of brucellosis and the level of anthropurgic enzooty of territories. In 2022, 467 cases of brucellosis were reported (0.32 per 100 000 of the population), which is 42.8 % higher than annual average values over 10 years. In 2023, a measure of stability of incidence rates, by 20–25 % above average long-term values, is to be expected. Incidence of brucellosis in humans will be within the range of 380–410 cases (0.26–0.28 per 100 000 of the population). The results of genotyping of B. melitensis strains point to an increase in the proportion of isolates with an MLVA-profile characteristic of strains from enzootic as regards brucellosis countries of Middle East and North Africa over past 20–25 years, which can indicate importation (introduction) of the infection from these territories to Russia through small ruminants and/or biomaterial from them

Software Solutions for Indication and Identification of Pathogenic Microoranisms Using Time-of-Flight Mass Spectrometry

The effectiveness of differentiation of bacterial pathogens using MALDI-TOF mass spectrometry depends on the quality of sample preparation, compliance with mass spectrometric analysis parameters and statistical approaches used, implemented by various modern software tools. The review provides a brief description of the most known software used in the processing and bioinformation analysis of time-of-flight mass spectrometry data. A list of computer platforms, programs and environments, both commercial and publicly available, is presented. The results of indication and identification of pathogens of particularly dangerous and natural-focal infections by MALDI-TOF mass spectrometry using publicly available software – programming language R, Mass-Up, MicrobeMS, licensed – MatLab, ClinProTools, as well as free web applications, including, Speclust, Ribopeaksare provided. The data on usage of such well-known platforms as MALDI BioTyper, SARAMIS Vitek-MS and Andromas (Andromas SAS, France) for inter- and intra-specific differentiation of closely related species are presented. Results of identification and differentiation of microorganisms applying MALDI-TOF mass spectrometry based on detection of specific proteins for cross-comparison – biomarkers – are given. The analysis shows that the programming language R environment is one of the publicly available universal platforms with an optimal combination of algorithms for processing and interpreting of a large array of mass spectrometric data

STUDY OF THE MASS-SPECTORS’ FEATURES OF SPORES AND VEGETATIVE FORMS OF BACILLUS ANTHRACIS BY THE METHOD OF TIME OF FLIGHT MASS-SPECTROMETRY

Aim. Investigation of the features of the protein profiles of the spore and vegetative form of the anthrax microbe by the MALDI-TOF MS method using the resources of the Mass-Up program and the package for the statistical software with open source code R. Materials and methods. Spores and vegetative forms of 32 strains of B. anthracis from the collection of microorganisms of the Stavropol Research Institute for Plague Control, including 8 strains isolated from an outbreak of anthrax in Yamal in 2016. Protein profiling was carried out on the Microflex MALDI-TOF mass spectrometer «Bruker Daltonics». Results. The alternative bioinformational-statistical approach used to analyze the MALDI-TOF mass spectra of the causative agent of anthrax made it possible to differentiate the spores and vegetative forms of the microbe based on the identification of the corresponding groups of biomarkers. Conclusion. A comparison of vegetative and spore cells of typical and atypical strains of anthrax causative agent on the basis of MALDI-TOF MS data was made. It has been experimentally confirmed that the protein profiles of cultures of Bacillus anthracis of the spore and vegetative form differ significantly, and this difference can be used to search for potential markers of each of the forms

Complex analysis of information technologies integration and adoption into medical organizations

Nowadays Information Technologies (IT) are taking more and more stable position in the process of work organization in various companies, and particularly among medical organizations. Present-day scientific society has already begun to research the topic of IT integration particularities into the Health organizations. Current article is focused on this issue. Our team has planned and provided a systematic review to understand which barriers can appear in different medical centers during implementation of the innovative IT into working process. The review has been fulfilled according to the designed protocol based on the actual recommendations of Preferred Reporting Items for Systematic Reviews and Meta-Analyses: The PRISMA Statement. In the course of the workflow we have explored 2050 articles complied with the requirements. As a result, 23 key publications have been chosen and analyzed in detail. After considering the list of issues and barriers associated with the IT Implementation into hospitals and medical centers have been highlighted and described. We have classified them to notional clusters depending on the reasons of their appearance as well. In addition our team has defined the list of practical recommendations following which the result of the IT integration will be improved, but the risk of a negative outcome in the future can be eliminated

Mass spectrometry analysis of protein blood extracts of animals with experimental brucellos

Aim. The aim of the present research was to study the possibility of direct detection of the causative agent of brucellosis in a biomaterial under experimental conditions via the MALDI-TOF MS method using Mass-Up program resources and a set of packages for open-source statistical software R. Materials and methods. We used laboratory mice infected with the causative agents of Brucellosis (strains B. melitensis 548, B. abortus 544, B. suis 1330) as models. Protein profiling was performed on a MALDI-TOF Microflex «Bruker Daltonics» mass spectrometer. Results. The bioinformatic-statistical approach used for analyzing MALDI-TOF mass spectra allows to carry out a direct detection of Brucella in the biomaterial; besides, it is possible to determinate their species via the identification of a group of biomarkers. Conclusion. It was experimentally confirmed that the protein profiles of the blood extracts of infected animals contain 11 markers, including 6 genus specific for Brucella spp., which can be associated with Brucella infection