Enhanced Colorimetric Differentiation between Staphylococcus aureus and Pseudomonas aeruginosa Using a Shape-Encoded Sensor Hydrogel

Zhiyuan Jia, Lauren Gwynne, Adam C. Sedgwick, Mareike Müller, George T. Williams, Andrew Toby A. Jenkins, Tony D. James, Holger Schönherr

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Abstract

Herein, we demonstrate a combined fluorescent probe/shape-encoded hydrogel strategy for the fast, sensitive, and selective detection of bacterial species via their characteristic enzymes. A poly(vinyl alcohol) (PVA) hydrogel loaded with the fluorescent probe N,N′-(3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl)bis(2,2,3,3,3-pentafluoropropanamide) (ACS-HNE) was designed for the detection of elastase, an enzyme produced by Pseudomonas aeruginosa. Likewise, a chitosan-derived hydrogel was loaded with the fluorescent probe 4-methylumbelliferyl-α-d-glucopyranoside (MUD) by entrapment for the selective detection of α-glucosidase, an enzyme produced by Staphylococcus aureus. For an observation time of 60 min, limits of detection (LODs) of ≤20 nM for elastase and ≤30 pM for α-glucosidase were obtained, which in the latter case is 3 orders of magnitude better than related chitosan systems with covalently coupled substrate. To illustrate the potential utility of these highly sensitive sensor hydrogels as a simple point-of-care test system, shaped hydrogel slabs representing the letters P and S were manufactured to detect P. aeruginosa and S. aureus, respectively. These shapes were shown to provide an additional unique color code under UV illumination corresponding to the characteristic enzyme produced by the corresponding bacteria. This study shows potential for the future development of an effective and simple point-of-care test for the rapid identification of bacterial species that can be operated by nonspecialists.

Original languageEnglish
Pages (from-to)4398-4407
Number of pages10
JournalACS Applied Bio Materials
Volume3
Issue number7
Early online date12 Jun 2020
DOIs
Publication statusPublished - 20 Jul 2020

Funding

The authors thank the EPSRC and the University of Bath for funding. The work in Austin was supported by the National Institutes of Health and the Robert A. Welch Foundation. This work was also supported in part by grant MR/N0137941/1 for the GW4 BIOMED DTP awarded to the Universities of Bath, Bristol, Cardiff, and Exeter from the Medical Research Council (MRC)/UKRI. A.C.S. thanks the EPSRC for a studentship. T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award. This work was further supported by the European Research Council (ERC Grant No. 279202 to HS), the equality office of the University of Siegen, the Max-Buchner-Forschungsstiftung Dechema (MBFSt-Kennziffer: 3671), and the University of Siegen.

Keywords

  • bacteria detection
  • bacteria differentiation
  • biosensors
  • fluorescent probe
  • hydrogels

ASJC Scopus subject areas

  • Biomaterials
  • General Chemistry
  • Biomedical Engineering
  • Biochemistry, medical

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