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Rapid Multi-Omics for Bacterial Identification Using Flow Injection-Ion Mobility-Mass Spectrometry.

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Rapid gas-phase ion mobility-mass spectrometry (IM-MS) enables faster bacterial identification by analyzing lipids and metabolites. This multiomic approach improves species-level detection and antibiotic resistance profiling in pathogens.

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Area of Science:

  • Clinical microbiology
  • Analytical chemistry
  • Biochemistry

Background:

  • Mass spectrometry (MS) in clinical microbiology accelerates positive culture identification.
  • Current protein-based MS methods face challenges in species-level identification due to high homology.
  • Lipid and small molecule-based MS strategies show potential for species-level identification and phenotype detection, including antibiotic resistance.

Purpose of the Study:

  • To leverage rapid gas-phase ion mobility (IM) separations coupled to MS for simultaneous detection of lipids and metabolites in bacterial pathogens.
  • To utilize flow-injection (FI) coupled with IM-MS as an alternative to liquid chromatography (LC) for bacterial identification.
  • To assess the performance of FI-IM-MS for distinguishing high-concern ESKAPE pathogens and enabling rapid multiomic analysis.

Main Methods:

  • Employed rapid gas-phase ion mobility (IM) separations directly coupled to mass spectrometry (MS).
  • Utilized flow-injection (FI) instead of liquid chromatography (LC) to rely on the IM dimension for structural separation.
  • Performed head-to-head comparison of FI-IM-MS with LC-IM-MS for distinguishing 24 strains of ESKAPE pathogens.

Main Results:

  • FI-IM-MS demonstrated comparable performance to LC-IM-MS in distinguishing ESKAPE pathogen strains.
  • Overall analysis time was significantly reduced from 17 to 2 minutes per injection using FI-IM-MS.
  • The IM dimension exhibited excellent stability and reproducibility, allowing the use of extracted IM peak areas for quantification.
  • Key features for bacterial discrimination were consistent between FI-IM-MS and HILIC-IM-MS data sets.

Conclusions:

  • Mobility-enabled rapid multiomics offers a powerful approach for bacterial pathogen analysis.
  • FI-IM-MS provides a faster alternative to LC-based methods while maintaining identification capabilities.
  • This technology holds promise for detecting subtle strain-level differences and antibiotic resistance phenotypes by incorporating diverse biomolecules.