# Differentiating bacteria by their unique surface interactions

**Authors:** Nicholas K. Kotoulas, Stephanie Tan, Justin R. Nodwell, M. Cynthia Goh

PMC · DOI: 10.1371/journal.pone.0327489 · 2025-06-30

## TL;DR

This paper introduces a new method to identify bacteria based on how they interact with different surfaces, which could help in diagnosing and managing antibiotic resistance.

## Contribution

The novel contribution is the development of surface interaction profiles (SIPs) for rapid and accurate bacterial differentiation.

## Key findings

- Surface interaction profiles (SIPs) successfully distinguished various pathogenic bacteria by Gram stain and species.
- SIPs identified antibiotic-resistant mutants like MRSA and VISA with high accuracy.
- The method was validated using blind tests and showed strong diagnostic potential.

## Abstract

New, rapid, and accessible approaches to bacterial detection are necessary to help curb the rising impacts of antimicrobial resistance. In this study, we introduce a technique that distinguishes bacteria through their unique surface interactions. By measuring and combining the interaction strengths of a bacterium across a set of chemically defined surfaces, we produced a novel bacterial identifier termed the surface interaction profile (SIP). The interaction strengths of twelve test bacteria across three discrete polyelectrolyte multilayer films (PEMs) were measured, facilitated by introducing each bacterial suspension to individual PEMs in microfluidic channels over a 10-minute interaction period and rinsing to remove bulk and loosely bound bacteria. The remaining surface-bound cells were counted via microscopy and plotted against suspension concentrations to build attachment curves whose slopes were measured as the strength of interaction for a given bacteria-PEM combination. These slopes were collected, per bacterial type, to produce each SIP. SIPs were capable of distinguishing between our pathogenic strains (Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterococcus faecalis, methicillin-resistant Staphylococcus aureus, and vancomycin-intermediate Staphylococcus aureus) by Gram stain and individual species, and each blind test pathogen was successfully identified through SIP comparison. Furthermore, SIPs were also successful at differentiating between select Staphylococcus aureus walKR mutants impacting cell wall metabolism and high-risk antibiotic resistance mutants (MRSA and VISA), highlighting the utility and future diagnostic potential of this technique.

## Linked entities

- **Species:** Klebsiella pneumoniae (taxon 573), Acinetobacter baumannii (taxon 470), Pseudomonas aeruginosa (taxon 287), Enterococcus faecalis (taxon 1351), Staphylococcus aureus (taxon 1280)

## Full-text entities

- **Chemicals:** methicillin (MESH:D008712), PEM (-), polyelectrolyte (MESH:D000071228), vancomycin (MESH:D014640)
- **Species:** Acinetobacter baumannii (species) [taxon 470], Enterococcus faecalis (species) [taxon 1351], Bacteria Latreille et al. 1825 (Bacteria stick insect, genus) [taxon 629395], Staphylococcus aureus (species) [taxon 1280], Klebsiella pneumoniae (species) [taxon 573], Pseudomonas aeruginosa (species) [taxon 287]

## Figures

3 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12208452/full.md

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Source: https://tomesphere.com/paper/PMC12208452