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Specific protease activity indicates the degree of Pseudomonas aeruginosa infection in chronic infected wounds

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Abstract

Chronic non-healing wounds are a major health problem with resident bacteria strongly implicated in their impaired healing. A rapid-screen to provide detailed knowledge of wound bacterial populations would therefore be of value and help prevent unnecessary and indiscriminate use of antibiotics—a process associated with promoting antibiotic resistance. We analysed chronic wound fluid samples, which had been assessed for microbial content, using 20 different fluorescent labelled peptide substrates to determine whether protease activity correlated with the bacterial load. Eight of the peptide substrates showed significant release of fluorescence after reaction with some of the wound samples. Comparison of wound fluid protease activities with the microbiological data indicated that there was no correlation between bacterial counts and enzyme activity for most of the substrates tested. However, two of the peptide substrates produced a signal corresponding with the microbial data revealing a strong positive correlation with Pseudomonas aeruginosa numbers. This demonstrated that short fluorescent labelled peptides can be used to detect protease activity in chronic wound fluid samples. The finding that two peptides were specific indicators for the presence of P. aeruginosa may be the basis for a diagnostic test to determine wound colonisation by this organism.

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Acknowledgements

This work was funded by the Engineering and Physical Sciences Research Council (EPSRC), grant number EP/D505445/1.

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Author notes

  1. P. E. Price

    Present address: School of Healthcare Studies, Cardiff University, Ty Dewi Sant, Heath Park, Cardiff, CF14 4XN, UK

Authors and Affiliations

  1. Pharmaceutical Science Division, Analytical Sciences Research Group, School of Biomedical & Health Sciences, King’s College London, Franklin-Wilkins Building, 150 Stamford Street, London, SE1 9NH, UK

    D. Wildeboer, F. Jeganathan, R. A. Abuknesha & R. G. Price

  2. School of Dentistry, Tissue Engineering & Reparative Dentistry, Cardiff University, Heath Park, Cardiff, CF14 4XY, UK

    K. E. Hill, D. W. Williams, A. D. Riddell, D. W. Thomas & P. Stephens

  3. School of Medicine, Department of Wound Healing, Cardiff University, Heath Park, Cardiff, CF14 4XN, UK

    A. D. Riddell & P. E. Price

  4. Natural Science Department, School of Health and Social Science, Middlesex University, The Burroughs, London, NW4 4BT, UK

    D. Wildeboer

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  1. D. Wildeboer
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  2. K. E. Hill
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Correspondence to D. Wildeboer.

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Wildeboer, D., Hill, K.E., Jeganathan, F. et al. Specific protease activity indicates the degree of Pseudomonas aeruginosa infection in chronic infected wounds. Eur J Clin Microbiol Infect Dis 31, 2183–2189 (2012). https://doi.org/10.1007/s10096-012-1553-6

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  • DOI: https://doi.org/10.1007/s10096-012-1553-6

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