scholarly journals Proteomic profiling of Yersinia pestis strains circulating in the area of natural plague foci of North Caucasus and Transcaucasia

2019 ◽  
pp. 18-25
Author(s):  
E. A. Koteneva ◽  
E. S. Kotenev ◽  
A. V. Kalinin ◽  
N. S. Tsarеva ◽  
L. A. Kot ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Yersinia Pestis ◽  
Mass Spectra ◽  
Molecular Genetic ◽  
Proteomic Profiling ◽  
North Caucasus ◽  
Biochemical Tests ◽  
Maldi Tof ◽  
Epizootic Process ◽  
Characteristic Features

Aim. To create a database of mass spectra of Yersinia pestis strains, which will differentiate the strains of the main and Caucasian subspecies of the plague agent by MALDI-TOF MS. Materials and methods. MALDITOF mass spectrometry was used to study 50 strains of Y. pestis, isolated on the territory of 7 natural plague foci of the Caucasus and Transcaucasia in the period 1950-2012. The removal of mass spectra of extracts of cells of Y. pestis was performed using the mass spectrometer Microflex LT «Bruker Daltonics». The results were processed and analyzed in FlexAnalysis programs, аnd MALDI Biotyper V. 3.0. Results. Тhis study showed that mass spectra have characteristic features that allow differentiating strains of the main (Y. pestis pestis) and subspecies (Y. pestis caucasica). Peaks characteristic of each subspecies were detected. The presence in Y. pestis caucasica subspecies peaks characteristic of the ancestral form — Y. pseudotuberculosis indicates the ancient origin of this group, which is consistent with the data of molecular genetic and WGS analysis given in other publications. Conclusion. Тhis work shows the possibility of applying the MALDI-TOF method of mass spectrometry for rapid differentiation of strains of the main subspecies Y. pestis pestis from the subspecies Y. pestis caucasica, which have different significance in the development and maintenance of the epizootic process in natural plague foci as well as different virulence for humans. Identification of the strain to the subspecies level requires carrying out culture and biochemical tests, which can take several days. The proposed method makes it possible to differentiate and obtain a result within half an hour after receiving a pure culture.

scholarly journals Application of MALDI-TOF mass spectrometry in clinical diagnostic microbiology

2014 ◽  
Vol 8 (09) ◽  
pp. 1081-1088 ◽  
Author(s):  
Elena De Carolis ◽  
Antonietta Vella ◽  
Luisa Vaccaro ◽  
Riccardo Torelli ◽  
Teresa Spanu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Drug Susceptibility ◽  
Biochemical Tests ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Bacteria And Fungi ◽  
Diagnostic Microbiology ◽  
Tof Ms

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently emerged as a powerful technique for identification of microorganisms, changing the workflow of well-established laboratories so that its impact on microbiological diagnostics has been unparalleled. In comparison with conventional identification methods that rely on biochemical tests and require long incubation procedures, MALDI-TOF MS has the advantage of identifying bacteria and fungi directly from colonies grown on culture plates in a few minutes and with simple procedures. Numerous studies on different systems available demonstrate the reliability and accuracy of the method, and new frontiers have been explored besides microbial species level identification, such as direct identification of pathogens from positive blood cultures, subtyping, and drug susceptibility detection.

scholarly journals Characterization of aliphatic hyperbranched polyesters by MALDI-TOF mass spectrometry

2007 ◽  
Vol 61 (6) ◽  
pp. 333-341
Author(s):  
Jasna Vukovic ◽  
Slobodan Jovanovic ◽  
Manfred Lechner
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectra ◽  
Degree Of Polymerization ◽  
Side Reactions ◽  
Maldi Tof Ms ◽  
Hyperbranched Polyesters ◽  
Maldi Tof ◽  
Maldi Tof Mass Spectra ◽  
Tof Ms

In this work, MALDI-TOF mass spectrometry was used for the characterization of aliphatic hyperbranched polyesters (AHBP), synthesized from 2,2-bis(hydroxymethyl)propionic acid (bis-MPA) and di-trimethylolpropane. From the obtained results it was concluded that it was not possible to take complete advantages of MALDI-TOF MS in this particular case, since the AHBP used in this work were polydisperse. The intensity of the signals from the high mass tail of these samples (pseudo generation higher than four) was underestimated and insufficient to distinguish it from the baseline and to use it for the analysis of the spectra. As a consequence of that, lower values of the Mn were obtained. At the same time, Mw were also underestimated, which led to very low values of the polydispersity index. On the other hand, it was possible to obtain molar masses of individual molecules from the MALDI-TOF mass spectra of AHBP and to qualitatively determine the extent of cyclization (side reactions) at each degree of polymerization. Using the adequate set of equations and results obtained from MALDI-TOF mass spectra of AHBP, every signal from the spectra was identified. The obtained results show that formation of poly(bis-MPA), intramolecular esterification and intramolecular etherification occurred as side reactions during the synthesis of these polyesters. The relative amount of the cycles increases with the number of pseudo generation (from the second up to the fifth pseudo generation). It was also observed that the relative proportion of the signals which represent cyclic structures increases with the increasing degree of polymerization. In this work the basic principles of MALDI-TOF MS are also presented, as well as, a review of adequate published articles.

scholarly journals Understanding the chemical basis for the preferential ionization of specific biomolecules in mass spectrometry analysis of microbial cells

2019 ◽  
Author(s):  
Wenfa Ng
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectra ◽  
Mass Spectrometry Analysis ◽  
Chemical Pretreatment ◽  
Maldi Tof Ms ◽  
Spectrometry Analysis ◽  
Chemical Basis ◽  
Maldi Tof ◽  
Different Types ◽  
Tof Ms

Mass spectrometry-enabled microbial identification has successfully demonstrated the feasibility of using profiled biomolecules for identifying microorganisms based on a chemometric or proteome database search approach. However, mechanisms driving the preferential ionization and detection of particular biomolecules in various types of mass spectrometry remain poorly understood. Specifically, mass spectra obtained from different microbial species remain poorly annotated with respect to the specific types of biomolecules accounting for the peaks. For example, while ribosomal proteins are known to be a significant class of biomolecules that could partially account for the profiled mass peaks in mass spectra of microorganisms, other classes of proteins and biomolecules remain poorly annotated. This raises the important question of how different mass spectrometry approaches ionize different types of biomolecules from a cellular matrix. Specifically, mass spectra of microorganisms reveal that only a couple of mass peaks could capture the phylogeny of a species. However, the proteome of a cell is much larger and more complicated, and yet is not fully profiled by different types of mass spectrometry methods. For example, electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) could only provide a small snapshot of the entire bacterial proteome. It could be argued that different mass spectrometry methods provide complementary views of a particular proteome. However, the question remains, how do proteins and biomolecules interact with the different sample preparation and mass spectrometry analysis methods for generating an ion cloud for separation in a mass spectrometer? Thus, efforts could be directed towards understanding how different types of proteins could be preferentially ionized by MALDI-TOF MS. Specifically, different reagents could be used to perform chemical pretreatment on the proteome, which would subsequently be analyzed by mass spectrometry. Thus, a correlative map between types of chemical pretreatment used and the corresponding mass spectra could be obtained. Collectively, knowledge gleaned from the research would illuminate the chemical basis by which specific biomolecules are preferentially ionized under particular conditions, which would inform the development of strategies for increasing the subset of biomolecules ionized from a cellular proteome. Such chemical rules would also aid in the interpretation of mass spectra obtained, particularly in understanding the biological context of the experiment. Overall, the key goal of this research is to help answer the question: what is the biological basis and context of the mass spectrum obtained from cells?

Determination of sensitivity of microorganisms of the Klebsiella genus to antimicrobial drugs by the MALDI-TOF MS method

Bacteriology ◽  
2020 ◽  
Vol 5 (3) ◽  
pp. 8-13
Author(s):  
A.A. Samoilova ◽  
◽  
I.V. Likhachev ◽  
E.V. Zueva ◽  
E.V. Rogacheva ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Antibiotic Resistance ◽  
Mass Spectra ◽  
Antimicrobial Agents ◽  
Antimicrobial Drugs ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Resistance Markers ◽  
Klebsiella Spp

Actual microbiological diagnostics of infections caused by Klebsiella spp. should include isolation of the strain, its identification and fastest possible determination of the pathogen susceptibility to antimicrobial agents. We evaluated the prospects of using MALDI-TOF mass spectrometry to determine the susceptibility of Klebsiella spp. strains to antimicrobial agents. According to the results of mass spectrometry Klebsiella spp. strains analysis, we carried out cluster analysis by the UPGMA method based on mass spectra and data on the susceptibility of the studied strains to antimicrobial agents, and then studied the obtained dendrograms. We identified the areas with the highest probability of the location of antibiotic resistance markers by comparing the mass spectra of susceptible and resistant microorganisms at different concentrations of antimicrobial agents. Thus, using MALDI-TOF mass spectrometry, a new direction in assessing the susceptibility of Klebsiella spp. to antimicrobial agents can be formed. Key words: antibiotic resistance, Klebsiella spp., MALDI-TOF mass spectrometry

scholarly journals A novel and rapid approach to yeast differentiation using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry

2003 ◽  
Vol 17 (1) ◽  
pp. 31-38 ◽  
Author(s):  
Richard E. Sherburn ◽  
Richard O. Jenkins
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectra ◽  
Time Of Flight ◽  
Laser Desorption ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Tof Ms ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

Matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF-MS) was investigated as a method for the rapid identification of yeast cells. Following pretreatment of yeast samples with a cell wall digesting enzyme (lyticase), distinct and reproducible mass spectra over them/zrange 2,000 to 16,000 were obtained by MALDI-TOF-MS. Using an optimised procedure, characteristic mass spectra that distinguished between Candida spp. and between strains of Saccharomyces cerevisiae were produced. The approach offers the potential for rapid differentiation of yeasts in clinical diagnosis and in the fermentation industries.

scholarly journals Discrimination of Bacillus cereus Group Members by MALDI-TOF Mass Spectrometry

2021 ◽  
Vol 9 (6) ◽  
pp. 1202
Author(s):  
Viviana Manzulli ◽  
Valeria Rondinone ◽  
Alessandro Buchicchio ◽  
Luigina Serrecchia ◽  
Dora Cipolletta ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bacillus Cereus ◽  
Mass Spectra ◽  
Critical Time ◽  
Principal Component ◽  
Etiologic Agent ◽  
Bacillus Cereus Group ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

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 Bacillus cereus group (57 B. anthracis, 49 B. cereus, 1 B. mycoides, 18 B. wiedmannii, 27 B. thuringiensis, 7 B. toyonensis and 1 B. weihenstephanensis) 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 B. toyonensis and B. wiedmannii 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 B. cereus group and to make a reliable identification of Bacillus anthracis, 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.

scholarly journals Understanding the chemical basis for the preferential ionization of specific biomolecules in mass spectrometry analysis of microbial cells

2019 ◽  
Author(s):  
Wenfa Ng
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectra ◽  
Mass Spectrometry Analysis ◽  
Chemical Pretreatment ◽  
Maldi Tof Ms ◽  
Spectrometry Analysis ◽  
Chemical Basis ◽  
Maldi Tof ◽  
Different Types ◽  
Tof Ms

Mass spectrometry-enabled microbial identification has successfully demonstrated the feasibility of using profiled biomolecules for identifying microorganisms based on a chemometric or proteome database search approach. However, mechanisms driving the preferential ionization and detection of particular biomolecules in various types of mass spectrometry remain poorly understood. Specifically, mass spectra obtained from different microbial species remain poorly annotated with respect to the specific types of biomolecules accounting for the peaks. For example, while ribosomal proteins are known to be a significant class of biomolecules that could partially account for the profiled mass peaks in mass spectra of microorganisms, other classes of proteins and biomolecules remain poorly annotated. This raises the important question of how different mass spectrometry approaches ionize different types of biomolecules from a cellular matrix. Specifically, mass spectra of microorganisms reveal that only a couple of mass peaks could capture the phylogeny of a species. However, the proteome of a cell is much larger and more complicated, and yet is not fully profiled by different types of mass spectrometry methods. For example, electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) could only provide a small snapshot of the entire bacterial proteome. It could be argued that different mass spectrometry methods provide complementary views of a particular proteome. However, the question remains, how do proteins and biomolecules interact with the different sample preparation and mass spectrometry analysis methods for generating an ion cloud for separation in a mass spectrometer? Thus, efforts could be directed towards understanding how different types of proteins could be preferentially ionized by MALDI-TOF MS. Specifically, different reagents could be used to perform chemical pretreatment on the proteome, which would subsequently be analyzed by mass spectrometry. Thus, a correlative map between types of chemical pretreatment used and the corresponding mass spectra could be obtained. Collectively, knowledge gleaned from the research would illuminate the chemical basis by which specific biomolecules are preferentially ionized under particular conditions, which would inform the development of strategies for increasing the subset of biomolecules ionized from a cellular proteome. Such chemical rules would also aid in the interpretation of mass spectra obtained, particularly in understanding the biological context of the experiment. Overall, the key goal of this research is to help answer the question: what is the biological basis and context of the mass spectrum obtained from cells?

scholarly journals A workflow to integrate pre-processing, analysis and comparison of MALDI-ToF mass spectra in GeenaR

2016 ◽  
Author(s):  
Eugenio Del Prete ◽  
Angelo Facchiano ◽  
Aldo Profumo ◽  
Claudia Angelini ◽  
Paolo Romano
Keyword(s):  
Mass Spectrometry ◽  
Statistical Methods ◽  
Mass Spectra ◽  
Large Scale ◽  
Heuristic Algorithms ◽  
Single Spectrum ◽  
Spectra Analysis ◽  
Maldi Tof ◽  
Maldi Tof Mass Spectra ◽  
R Packages

Many large-scale proteomics studies have been performed in the last years, and this field of investigation is expanding up. If the analysis of any single spectrum can be performed by tools already made available along with the mass spectrometry (MS) instrumentation, comparison of spectra on a large scale represents a complex aspect of the analysis and interpretation of the study. Recently, we developed Geena 2, a tool for the automation of different steps in the MALDI/ToF MS data analysis. Integration of further tools can be performed, in order to improve some aspects of the whole workflow: the input of more data formats, the implementation of new algorithms for data cleaning, the graphical visualization and the reporting of the results, the use of advanced statistics for the comparison of mass spectra. For this motivations, we are now developing GeenaR, a new robust web tool for pre-processing, analysing, visualizing and comparing a set of MALDI-ToF mass spectra. The aim of this work is the presentation of on-going developments. The first results will be presented at the conference. GeenaR is being written in PHP, Perl (from Geena 2) and R languages. The R packages used are MALDIquant and MALDIquantForeign for mass spectra pre-processing and analysis, OrgMassSpecR for mass spectra comparison, dendextend and pvclust for clustering, and sda and crossval for variable selection. The system is being implemented in a LAMP (Linux, Apache, MySQL, PHP) environment. Proper interfaces between PHP on one side and perl and R on the other are then implemented. The aim of GeenaR is to provide to the users a wider range of statistical methods and graphical results, without making it more difficult to use for researchers with little expertise in programming. In order to achieve this goal, we have taken advantage of the availability of several packages, written in R language, for mass spectrometry statistical that are going to be integrated in the system. The complete pipeline of GeenaR includes some features already available in Geena 2 plus others under development thanks to the integration of the R environment. In fact, an original set of heuristic algorithms is already available in Geena 2. In particular, they are the identification of isotopic peaks by taking into account molecular weight of signals and the related trend of abundances; the normalization on the basis of a reference standard molecule; the peak selection by means of a threshold line, built by linearly interpolating values provided for given m/z values; the alignment, by selecting the nearest peaks, within a limited m/z difference, in the different mass spectra. By means of some R packages, GeenaR adds new statistical methods which are highly relevant for mass spectra analysis. (Abstract truncated at 3,000 characters - the full version is available in the pdf file)

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