Electrochemical detection of Pseudomonas in wound exudate samples from patients with chronic wounds

Wound Repair Regen. 2016 Mar;24(2):366-72. doi: 10.1111/wrr.12414. Epub 2016 Mar 6.

Abstract

In clinical practice, point-of-care diagnostic testing has progressed rapidly in the last decade. For the field of wound care, there is a compelling need to develop rapid alternatives for bacterial identification in the clinical setting, where it generally takes over 24 hours to receive a positive identification. Even new molecular and biochemical identification methods require an initial incubation period of several hours to obtain a sufficient number of cells prior to performing the analysis. Here we report the use of an inexpensive, disposable electrochemical sensor to detect pyocyanin, a unique, redox-active quorum sensing molecule released by Pseudomonas aeruginosa, in wound fluid from patients with chronic wounds enrolled in the WE-HEAL Study. By measuring the metabolite excreted by the cells, this electrochemical detection strategy eliminates sample preparation, takes less than a minute to complete, and requires only 7.5 μL of sample to complete the analysis. The electrochemical results were compared against 16S rRNA profiling using 454 pyrosequencing. Blind identification yielded 9 correct matches, 2 false negatives, and 3 false positives giving a sensitivity of 71% and specificity of 57% for detection of Pseudomonas. Ongoing enhancement and development of this approach with a view to develop a rapid point-of-care diagnostic tool is planned.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adult
  • Biofilms / growth & development
  • Biosensing Techniques* / instrumentation
  • Biosensing Techniques* / trends
  • Chronic Disease
  • Disposable Equipment
  • Electrochemical Techniques* / instrumentation
  • Electrochemical Techniques* / trends
  • Equipment Design
  • Exudates and Transudates / microbiology*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Point-of-Care Systems* / trends
  • Pseudomonas Infections / microbiology*
  • Pyocyanine / analysis
  • RNA, Ribosomal, 16S / analysis
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Time Factors
  • Wound Infection / microbiology*

Substances

  • RNA, Ribosomal, 16S
  • Pyocyanine