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Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens
Microbes have diverse metabolic capabilities and differences in these phenotypes are critical for differentiating strains, species, and broader taxa of microorganisms. Recent advances in liquid chromatography-mass spectrometry (LC-MS) allow researchers to track the complex combinations of molecules...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9354526/ https://www.ncbi.nlm.nih.gov/pubmed/35935218 http://dx.doi.org/10.3389/fmicb.2022.957158 |
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author | Mapar, Maryam Rydzak, Thomas Groves, Ryan A. Lewis, Ian A. |
author_facet | Mapar, Maryam Rydzak, Thomas Groves, Ryan A. Lewis, Ian A. |
author_sort | Mapar, Maryam |
collection | PubMed |
description | Microbes have diverse metabolic capabilities and differences in these phenotypes are critical for differentiating strains, species, and broader taxa of microorganisms. Recent advances in liquid chromatography-mass spectrometry (LC-MS) allow researchers to track the complex combinations of molecules that are taken up by each cell type and to quantify the rates that individual metabolites enter or exit the cells. This metabolomics-based approach allows complex metabolic phenotypes to be captured in a single assay, enables computational models of microbial metabolism to be constructed, and can serve as a diagnostic approach for clinical microbiology. Unfortunately, metabolic phenotypes are directly affected by the molecular composition of the culture medium and many traditional media are subject to molecular-level heterogeneity. Herein, we show that commercially sourced Mueller Hinton (MH) medium, a Clinical and Laboratory Standards Institute (CLSI) approved medium for clinical microbiology, has significant lot-to-lot and supplier-to-supplier variability in the concentrations of individual nutrients. We show that this variability does not affect microbial growth rates but does affect the metabolic phenotypes observed in vitro—including metabolic phenotypes that distinguish six common pathogens. To address this, we used a combination of isotope-labeling, substrate exclusion, and nutritional supplementation experiments using Roswell Park Memorial Institute (RPMI) medium to identify the specific nutrients used by the microbes to produce diagnostic biomarkers, and to formulate a Biomarker Enrichment Medium (BEM) as an alternative to complex undefined media for metabolomics research, clinical diagnostics, antibiotic susceptibility testing, and other applications where the analysis of stable microbial metabolic phenotypes is important. |
format | Online Article Text |
id | pubmed-9354526 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-93545262022-08-06 Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens Mapar, Maryam Rydzak, Thomas Groves, Ryan A. Lewis, Ian A. Front Microbiol Microbiology Microbes have diverse metabolic capabilities and differences in these phenotypes are critical for differentiating strains, species, and broader taxa of microorganisms. Recent advances in liquid chromatography-mass spectrometry (LC-MS) allow researchers to track the complex combinations of molecules that are taken up by each cell type and to quantify the rates that individual metabolites enter or exit the cells. This metabolomics-based approach allows complex metabolic phenotypes to be captured in a single assay, enables computational models of microbial metabolism to be constructed, and can serve as a diagnostic approach for clinical microbiology. Unfortunately, metabolic phenotypes are directly affected by the molecular composition of the culture medium and many traditional media are subject to molecular-level heterogeneity. Herein, we show that commercially sourced Mueller Hinton (MH) medium, a Clinical and Laboratory Standards Institute (CLSI) approved medium for clinical microbiology, has significant lot-to-lot and supplier-to-supplier variability in the concentrations of individual nutrients. We show that this variability does not affect microbial growth rates but does affect the metabolic phenotypes observed in vitro—including metabolic phenotypes that distinguish six common pathogens. To address this, we used a combination of isotope-labeling, substrate exclusion, and nutritional supplementation experiments using Roswell Park Memorial Institute (RPMI) medium to identify the specific nutrients used by the microbes to produce diagnostic biomarkers, and to formulate a Biomarker Enrichment Medium (BEM) as an alternative to complex undefined media for metabolomics research, clinical diagnostics, antibiotic susceptibility testing, and other applications where the analysis of stable microbial metabolic phenotypes is important. Frontiers Media S.A. 2022-07-22 /pmc/articles/PMC9354526/ /pubmed/35935218 http://dx.doi.org/10.3389/fmicb.2022.957158 Text en Copyright © 2022 Mapar, Rydzak, Groves and Lewis. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Mapar, Maryam Rydzak, Thomas Groves, Ryan A. Lewis, Ian A. Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens |
title | Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens |
title_full | Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens |
title_fullStr | Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens |
title_full_unstemmed | Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens |
title_short | Biomarker enrichment medium: A defined medium for metabolomic analysis of microbial pathogens |
title_sort | biomarker enrichment medium: a defined medium for metabolomic analysis of microbial pathogens |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9354526/ https://www.ncbi.nlm.nih.gov/pubmed/35935218 http://dx.doi.org/10.3389/fmicb.2022.957158 |
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