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Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of microbial mixtures: a first impression of its applicability for the analysis of gastro-intestinal microbiota

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Abstract
Introduction & Objectives: Although scientists are more than ever conscious of the importance of the gastrointestinal (GI) microbiota and its influence on the health of the host, to date, our knowledge on the subject is still restricted. Despite the introduction of molecular culture-independent techniques, which already signified an important step forwards in a better understanding of the microbial composition of the GI tract, still a large part remains uncharted. In addition, there is also a need for applications which are less labor intensive and much faster. A technique recently emerging in the field of microbiology is matrix-assisted laser desorption ionization - time of flight mass spectrometry (MALDI-TOF MS). Its high sensitivity and high throughput made it a very interesting method for the characterization and identification of large amounts of microbial isolates. The fact that a given species can be identified through unique biomarker(s) (i.e. mass peaks with a certain m/z ratio in the MS spectrum) opens up new perspectives to be able to discriminate and identify different members of a mixed microbial community. With this, important advantages are the comparative simplicity of the mass spectra for a given species and the high reproducibility of the technique. In this study, the effect of an increasing complexity of a microbial mixture on the quality of the MS profiles and as a consequence the usability of MALDI-TOF for the analysis of more complex microbial communities such as those of the GI tract, was evaluated. In addition, an attempt was made to determine the sensitivity of the technique. Material & Methods: Mixtures of different species composition and/or increasing complexity were subjected to MALDI-TOF MS analysis. In parallel, denaturing gradient gel electrophoresis (DGGE), a commonly applied approach which has proven its usefulness studying GI microbiota, was carried out. Visual and cluster analysis were performed for data interpretation. Results & Discussion: Complexity of MS profiles increased with growing numbers of species added to the mixture. However, spectral profiles remained of good quality allowing to compare mixtures of different species composition. Despite the quality profiles, visual interpretation became more difficult as complexity increased. Current software limitations could not give an answer to this problem. As a result, individual members could not always be retrieved from the community profile, explaining the need for specialized data interpretation software allowing more in-depth analyses. Sensitivity of MALDI-TOF MS was at least as high as for DGGE with a detection limit situated around 1%. However, actual sensitivity could be higher as this observation could also be biased by the software and visual restrictions. Preliminary results, although based on strongly simplified model, indicate possibilities for MALDI-TOF MS as a fast and high throughput application for the analysis of more complex microbial communities like the GI microbiota.

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MLA
Van Hoorde, Koenraad, Peter Vandamme, and Anita Van Landschoot. “Matrix-assisted Laser Desorption Ionization-time of Flight Mass Spectrometry (MALDI-TOF MS) of Microbial Mixtures: a First Impression of Its Applicability for the Analysis of Gastro-intestinal Microbiota.” Gastro-Intestinal Microbial Ecology, Conference Abstracts. 2010. Print.
APA
Van Hoorde, Koenraad, Vandamme, P., & Van Landschoot, A. (2010). Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of microbial mixtures: a first impression of its applicability for the analysis of gastro-intestinal microbiota. Gastro-Intestinal Microbial Ecology, Conference abstracts. Presented at the 2010 International scientific conference on Gastro-Intestinal Microbial Ecology (GME 2010).
Chicago author-date
Van Hoorde, Koenraad, Peter Vandamme, and Anita Van Landschoot. 2010. “Matrix-assisted Laser Desorption Ionization-time of Flight Mass Spectrometry (MALDI-TOF MS) of Microbial Mixtures: a First Impression of Its Applicability for the Analysis of Gastro-intestinal Microbiota.” In Gastro-Intestinal Microbial Ecology, Conference Abstracts.
Chicago author-date (all authors)
Van Hoorde, Koenraad, Peter Vandamme, and Anita Van Landschoot. 2010. “Matrix-assisted Laser Desorption Ionization-time of Flight Mass Spectrometry (MALDI-TOF MS) of Microbial Mixtures: a First Impression of Its Applicability for the Analysis of Gastro-intestinal Microbiota.” In Gastro-Intestinal Microbial Ecology, Conference Abstracts.
Vancouver
1.
Van Hoorde K, Vandamme P, Van Landschoot A. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of microbial mixtures: a first impression of its applicability for the analysis of gastro-intestinal microbiota. Gastro-Intestinal Microbial Ecology, Conference abstracts. 2010.
IEEE
[1]
K. Van Hoorde, P. Vandamme, and A. Van Landschoot, “Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of microbial mixtures: a first impression of its applicability for the analysis of gastro-intestinal microbiota,” in Gastro-Intestinal Microbial Ecology, Conference abstracts, Kosice, Slovakia, 2010.
@inproceedings{4274804,
  abstract     = {Introduction & Objectives: Although scientists are more than ever conscious of the importance of the gastrointestinal (GI) microbiota and its influence on the health of the host, to date, our knowledge on the subject is still restricted. Despite the introduction of molecular culture-independent techniques, which already signified an important step forwards in a better understanding of the microbial composition of the GI tract, still a large part remains uncharted.  In addition, there is also a need for applications which are less labor intensive and much faster. A technique recently emerging in the field of microbiology is matrix-assisted laser desorption ionization - time of flight mass spectrometry (MALDI-TOF MS). Its high sensitivity and high throughput made it a very interesting method for the characterization and identification of large amounts of microbial isolates. The fact that a given species can be identified through unique biomarker(s) (i.e. mass peaks with a certain m/z ratio in the MS spectrum) opens up new perspectives to be able to discriminate and identify different members of a mixed microbial community. With this, important advantages are the comparative simplicity of the mass spectra for a given species and the high reproducibility of the technique.
In this study, the effect of an increasing complexity of a microbial mixture on the quality of the MS profiles and as a consequence the usability of MALDI-TOF for the analysis of more complex microbial communities such as those of the GI tract, was evaluated. In addition, an attempt was made to determine the sensitivity of the technique.
Material & Methods: Mixtures of different species composition and/or increasing complexity were subjected to MALDI-TOF MS analysis. In parallel, denaturing gradient gel electrophoresis (DGGE),  a commonly applied approach which has proven its usefulness studying GI microbiota, was carried out. Visual and cluster analysis were performed for data interpretation.
Results & Discussion: Complexity of MS profiles increased with growing numbers of species added to the mixture. However, spectral profiles remained of good quality allowing to compare mixtures of different species composition. Despite the quality profiles, visual interpretation became more difficult as complexity increased. Current software limitations could not give an answer to this problem. As a result, individual members could not always be retrieved from the community profile, explaining the need for specialized data interpretation software allowing more in-depth analyses. Sensitivity of MALDI-TOF MS was at least as high as for DGGE with a detection limit situated around 1%. However, actual sensitivity could be higher as this observation could also be biased by the software and visual restrictions. Preliminary results, although based on strongly simplified model, indicate possibilities for MALDI-TOF MS as a fast and high throughput application for the analysis of more complex microbial communities like the GI microbiota.},
  author       = {Van Hoorde, Koenraad and Vandamme, Peter and Van Landschoot, Anita},
  booktitle    = {Gastro-Intestinal Microbial Ecology, Conference abstracts},
  language     = {eng},
  location     = {Kosice, Slovakia},
  title        = {Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of microbial mixtures: a first impression of its applicability for the analysis of gastro-intestinal microbiota},
  year         = {2010},
}