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Updated: Sep 18, 2025
06:34
Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
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Rapid Multi-Omics for Bacterial Identification Using Flow Injection-Ion Mobility-Mass Spectrometry.
Hannah M Hynds1, Jana M Carpenter1, Kelly M Hines1
1Department of Chemistry, University of Georgia, Athens, Georgia 30602, United States.
Analytical Chemistry
|June 24, 2025
Summary
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.
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.
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