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Mass Spectrometry-based PhyloProteomics (MSPP): A novel microbial typing Method.

Zautner AE, Masanta WO, Weig M, Groß U, Bader O - Sci Rep (2015)

Bottom Line: Performing comparably to laborious multilocus and whole genome sequence typing (wgMLST)-approaches it is feasible to build phyloproteomic dendrograms using hierarchical cluster analysis.MSPP bears a high potential especially for identification of chromosomal localised virulence or antimicrobial resistance factors associated with evolutionary relatedness.In this study the principle of MSPP-typing was demonstrated on a Campylobacter jejuni ssp. jejuni isolate collection and MSPP was compared to MLST.

View Article: PubMed Central - PubMed

Affiliation: Institut für Medizinische Mikrobiologie, Universitätsmedizin Göttingen, Kreuzbergring 57, 37075 Göttingen, Germany.

ABSTRACT
MALDI-TOF-MS of microorganisms, which identifies microbes based on masses of high abundant low molecular weight proteins, is rapidly advancing to become another standard method in clinical routine laboratory diagnostics. Allelic isoforms of these proteins result in varying masses of detectable biomarker ions. These variations give rise to a novel typing method for microorganisms named mass spectrometry-based phyloproteomics (MSPP). The base of MSPP is an amino acid sequence list of allelic isoforms caused by non-synonymous mutations in biomarker genes, which were detectable as mass shifts in an overlay of calibrated MALDI-TOF spectra. Thus, for each isolate a combination of amino acid sequences can be deduced from the scheme of recordable biomarker masses. Performing comparably to laborious multilocus and whole genome sequence typing (wgMLST)-approaches it is feasible to build phyloproteomic dendrograms using hierarchical cluster analysis. MSPP bears a high potential especially for identification of chromosomal localised virulence or antimicrobial resistance factors associated with evolutionary relatedness. In this study the principle of MSPP-typing was demonstrated on a Campylobacter jejuni ssp. jejuni isolate collection and MSPP was compared to MLST.

No MeSH data available.


Schematic overview of MSPP workflow.(a) Microbial culture of all isolates included in a current typing setting. (b) Recording of ICMS mass spectra with subsequent calibration, smoothing and baseline subtraction. (c) Detection and quantification of mass differences of biomarker ions due to different allelic isoforms in particular strains. (d) Database search for allelic isoforms that correspond to the detected mass differences. (e) Joining of amino acid sequences from all biomarker ions included in the MSPP scheme (f) Construction of taxonomic dendrograms using the UPGMA method.
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f2: Schematic overview of MSPP workflow.(a) Microbial culture of all isolates included in a current typing setting. (b) Recording of ICMS mass spectra with subsequent calibration, smoothing and baseline subtraction. (c) Detection and quantification of mass differences of biomarker ions due to different allelic isoforms in particular strains. (d) Database search for allelic isoforms that correspond to the detected mass differences. (e) Joining of amino acid sequences from all biomarker ions included in the MSPP scheme (f) Construction of taxonomic dendrograms using the UPGMA method.

Mentions: The MSPP typing procedure includes the following steps: (a) Culturing the microbial isolates to be typed, and (b) recording of ICMS spectra for each isolate (c) pre-processing and calibration of the recorded spectra, (d) measuring of mass shifts with reference to the genomic sequenced reference strain, (e) identification of particular allelic isoforms by matching the mass shifts with the database deduced isoform amino acid sequence set, (f) combination of amino acid sequences in the corresponding MSPP typing scheme and calculation of a phyloproteomic UPGMA-tree (Fig. 2).


Mass Spectrometry-based PhyloProteomics (MSPP): A novel microbial typing Method.

Zautner AE, Masanta WO, Weig M, Groß U, Bader O - Sci Rep (2015)

Schematic overview of MSPP workflow.(a) Microbial culture of all isolates included in a current typing setting. (b) Recording of ICMS mass spectra with subsequent calibration, smoothing and baseline subtraction. (c) Detection and quantification of mass differences of biomarker ions due to different allelic isoforms in particular strains. (d) Database search for allelic isoforms that correspond to the detected mass differences. (e) Joining of amino acid sequences from all biomarker ions included in the MSPP scheme (f) Construction of taxonomic dendrograms using the UPGMA method.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4548220&req=5

f2: Schematic overview of MSPP workflow.(a) Microbial culture of all isolates included in a current typing setting. (b) Recording of ICMS mass spectra with subsequent calibration, smoothing and baseline subtraction. (c) Detection and quantification of mass differences of biomarker ions due to different allelic isoforms in particular strains. (d) Database search for allelic isoforms that correspond to the detected mass differences. (e) Joining of amino acid sequences from all biomarker ions included in the MSPP scheme (f) Construction of taxonomic dendrograms using the UPGMA method.
Mentions: The MSPP typing procedure includes the following steps: (a) Culturing the microbial isolates to be typed, and (b) recording of ICMS spectra for each isolate (c) pre-processing and calibration of the recorded spectra, (d) measuring of mass shifts with reference to the genomic sequenced reference strain, (e) identification of particular allelic isoforms by matching the mass shifts with the database deduced isoform amino acid sequence set, (f) combination of amino acid sequences in the corresponding MSPP typing scheme and calculation of a phyloproteomic UPGMA-tree (Fig. 2).

Bottom Line: Performing comparably to laborious multilocus and whole genome sequence typing (wgMLST)-approaches it is feasible to build phyloproteomic dendrograms using hierarchical cluster analysis.MSPP bears a high potential especially for identification of chromosomal localised virulence or antimicrobial resistance factors associated with evolutionary relatedness.In this study the principle of MSPP-typing was demonstrated on a Campylobacter jejuni ssp. jejuni isolate collection and MSPP was compared to MLST.

View Article: PubMed Central - PubMed

Affiliation: Institut für Medizinische Mikrobiologie, Universitätsmedizin Göttingen, Kreuzbergring 57, 37075 Göttingen, Germany.

ABSTRACT
MALDI-TOF-MS of microorganisms, which identifies microbes based on masses of high abundant low molecular weight proteins, is rapidly advancing to become another standard method in clinical routine laboratory diagnostics. Allelic isoforms of these proteins result in varying masses of detectable biomarker ions. These variations give rise to a novel typing method for microorganisms named mass spectrometry-based phyloproteomics (MSPP). The base of MSPP is an amino acid sequence list of allelic isoforms caused by non-synonymous mutations in biomarker genes, which were detectable as mass shifts in an overlay of calibrated MALDI-TOF spectra. Thus, for each isolate a combination of amino acid sequences can be deduced from the scheme of recordable biomarker masses. Performing comparably to laborious multilocus and whole genome sequence typing (wgMLST)-approaches it is feasible to build phyloproteomic dendrograms using hierarchical cluster analysis. MSPP bears a high potential especially for identification of chromosomal localised virulence or antimicrobial resistance factors associated with evolutionary relatedness. In this study the principle of MSPP-typing was demonstrated on a Campylobacter jejuni ssp. jejuni isolate collection and MSPP was compared to MLST.

No MeSH data available.