From Identification to Insight: Making Full Use of the Diagnostic Potential of MS/MS Proteotyping in Clinical Microbiology Using Efficient Bioinformatics

peer-reviewed · Journal of Proteome Research · 2025

peer-reviewed · Journal of Proteome Research · 2025. Thilo Muth et al. Accurate and rapid pathogen identification is critical for modern diagnostics, driving the need for…
Date 2025-11-07
Type peer-reviewed
Venue Journal of Proteome Research
Publisher American Chemical Society (ACS)
Contribution review
DOI 10.1021/acs.jproteome.5c00733
Citations (OpenAlex) 3
Venue 2-year citedness 3.83

Abstract

Accurate and rapid pathogen identification is critical for modern diagnostics, driving the need for technologies that combine speed, specificity, and functional insight. In this context, mass spectrometry (MS)-based proteomics is rapidly emerging as a transformative approach in clinical microbiology, offering comprehensive characterization of microorganisms and direct identification of protein biomarkers relevant to infectious disease diagnostics. While MS1-based approaches such as MALDI-TOF Biotyper have revolutionized species-level microbial identification through rapid and cost-effective workflows, they remain fundamentally limited in resolving strain-level differences and in detecting antimicrobial resistance (AMR) determinants. In contrast, so-called proteotyping enables the identification of organisms and resistance-associated proteins based on peptide sequence information obtained by tandem mass spectrometry (MS/MS), most commonly using shotgun proteomics. However, the clinical utility of proteotyping depends on the availability of efficient and accurate bioinformatics tools capable of handling large databases, disambiguating shared peptide sequences, and providing taxonomic and functional assignments with clinical relevance. This perspective highlights the diagnostic potential of MS/MS proteotyping and argues that advances in bioinformatics pipelines are needed for moving from microbial identification to actionable clinical insight.

Authors

  1. Thilo Muth · Federal Institute for Materials Research and Testing (BAM), Max Planck Institute for Dynamics of Complex Technical Systems, Robert Koch Institute, glyXera GmbH
  2. Roger Karlsson
  3. Yi-Kuo Yu · National Institutes of Health
  4. Gelio Alves · National Institutes of Health

Methods and tools

  • MS/MS proteotyping in clinical microbiology (perspective): Perspective arguing that MS2-based proteotyping should replace MS1 profiling such as MALDI-TOF Biotyper for clinical pathogen identification, and surveying the bioinformatics that would make it practical. Covers database search, spectral library and de novo approaches together.

Seen in the charts

Back to the full map

Back to top