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Adapting a commercial sample extraction protocol for biosafety level 3/4 compatible plasma metabolomics analysis.

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  • 1Austere Environments Consortium for Enhanced Sepsis Outcomes, Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, MD, USA.

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Summary

This study adapted metabolomics assays for analyzing high-risk infections, finding that while some metabolites are reliably measured, others require careful consideration due to extraction differences. This improves host biomarker discovery for infectious diseases.

Keywords:
BiosafetyHigh-risk pathogensInfectious diseasesMetabolomicsSample extraction

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Area of Science:

  • Biochemistry
  • Clinical Diagnostics
  • Infectious Disease Research

Background:

  • Sepsis and severe infections critically impair military readiness and incur significant costs.
  • Early sepsis diagnosis is vital for patient outcomes but hindered by biomarker discovery challenges.
  • Current host biomarker discovery protocols often require pathogen inactivation, limiting metabolomics assay utility.

Purpose of the Study:

  • To evaluate the impact of a pathogen inactivation method (MPLEx) on a commercial metabolomics assay (AbsoluteIDQ p180).
  • To assess the suitability of adapted metabolomics protocols for high-risk infectious disease studies.
  • To identify differences in metabolite detection and extraction efficiency between methods.

Main Methods:

  • Plasma samples from a sepsis cohort were analyzed using the AbsoluteIDQ p180 protocol with and without MPLEx.
  • Analyte detection rates, concentrations, and extraction efficiencies were compared between the two methods.
  • The study focused on metabolites, proteins, and lipids relevant to host-pathogen interactions.

Main Results:

  • Agreement between methods varied by metabolite class; amino acids, glycerophospholipids, sphingolipids, and monosaccharides showed good correlation.
  • The MPLEx-p180 method reduced detection rates for biogenic amines and acylcarnitines due to higher sample dilution.
  • Differences in overall extraction efficiencies were observed between the unmodified and MPLEx-enhanced protocols.

Conclusions:

  • The study demonstrates the extended applicability of commercial metabolomics assays for high-risk infectious disease research.
  • Consideration of metabolite extraction efficiencies and detection rates is crucial when combining data from different protocols.
  • Findings inform strategies for host biomarker discovery in challenging infectious disease contexts.