title: Electrochemical detection of Pseudomonas in wound exudate samples from patients with chronic wounds.
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.
authors.affiliation: Department of Chemical Engineering, Northeastern University, Boston, Massachusetts.
authors.forename: Hunter J
authors.initials: HJ
authors.lastname: Sismaet
authors.affiliation: Division of Rheumatology, Ideas to Health Laboratory, The George Washington University, School of Medicine and Health Sciences, NW, Washington, DC.
authors.forename: Anirban
authors.initials: A
authors.lastname: Banerjee
authors.affiliation: Division of Rheumatology, Ideas to Health Laboratory, The George Washington University, School of Medicine and Health Sciences, NW, Washington, DC.
authors.forename: Sean
authors.initials: S
authors.lastname: McNish
authors.affiliation: The J. Craig Venter Institute, Rockville, Maryland.
authors.forename: Yongwook
authors.initials: Y
authors.lastname: Choi
authors.affiliation: The J. Craig Venter Institute, Rockville, Maryland.
authors.forename: Manolito
authors.initials: M
authors.lastname: Torralba
authors.affiliation: The J. Craig Venter Institute, Rockville, Maryland.
authors.forename: Sarah
authors.initials: S
authors.lastname: Lucas
authors.affiliation: The J. Craig Venter Institute, Rockville, Maryland.
authors.forename: Agnes
authors.initials: A
authors.lastname: Chan
authors.affiliation: Division of Rheumatology, Ideas to Health Laboratory, The George Washington University, School of Medicine and Health Sciences, NW, Washington, DC.
authors.forename: Victoria K
authors.initials: VK
authors.lastname: Shanmugam
authors.affiliation: Department of Chemical Engineering, Northeastern University, Boston, Massachusetts.
authors.forename: Edgar D
authors.initials: ED
authors.lastname: Goluch
mesh_terms: D000328:Adult; D018441:Biofilms / Q000254:growth & development; D015374:Biosensing Techniques / Q000295:instrumentation / Q000639:trends; D002908:Chronic Disease; D004209:Disposable Equipment; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000639:trends; D004867:Equipment Design; D005122:Exudates and Transudates / Q000382:microbiology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D019095:Point-of-Care Systems / Q000639:trends; D011552:Pseudomonas Infections / Q000382:microbiology*; D011710:Pyocyanine / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014946:Wound Infection / Q000382:microbiology*
chemical_list: D012336:RNA, Ribosomal, 16S; D011710:Pyocyanine
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