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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 ?L 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.

SUBMITTER: Sismaet HJ

PROVIDER: S-EPMC4853203 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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

Sismaet Hunter J HJ Banerjee Anirban A McNish Sean S Choi Yongwook Y Torralba Manolito M Lucas Sarah S Chan Agnes A Shanmugam Victoria K VK Goluch Edgar D ED

Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society 20160306 2

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. ...[more]

PMID: 26815644

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Project description:Burn wound sepsis is currently the main cause of morbidity and mortality after burn trauma. Infections by notorious pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus, and Acinetobacter baumannii impair patient recovery and can even lead to fatality. In this study, we investigated the effect of burn wound exudates (BWEs) on the virulence of those pathogens. BWEs were collected within 7 days after burn trauma from 5 burn patients. We first monitored their effect on pathogen growth. In contrast to A. baumannii and S. aureus, P. aeruginosa was the only pathogen able to grow within these human fluids. Expression of typical virulence factors such as pyocyanin and pyoverdine was even enhanced compared the levels seen with standard laboratory medium. A detailed chemical composition analysis of BWE was performed, which enabled us to determine the major components of BWE and underline the metabolic modifications induced by burn trauma. These data are essential for the development of an artificial medium mimicking the burn wound environment and the establishment of an in vitro system to analyze the initial steps of burn wound infections. IMPORTANCE Microbial infection of severe burn wounds is currently a major medical challenge. Of the infections by bacteria able to colonize such injuries, those by Pseudomonas aeruginosa are among the most severe, causing major delays in burn patient recovery or leading to fatal issues. In this study, we investigated the growth properties of several burn wound pathogens in biological fluids secreted from human burn wounds. We found that P. aeruginosa strains were able to proliferate but not those of the other pathogens tested. In addition, burn wound exudates (BWEs) stimulate the expression of virulence factors in P. aeruginosa. The chemical composition analysis of BWEs enabled us to determine the major components of these fluids. These data are essential for the development of an artificial medium mimicking the burn wound environment and for in vitro analysis of the initial step in the development of burn wound infections.

Project description:BackgroundIn civilian life, from 11 to 40% of patients suffer from chronic pain after receiving injuries. There are almost no data on chronic pain in patients with gunshot wounds, isolated clinical cases have been published. The purpose of our study is to determine the factors that can potentially affect the results of treatment of such patients, namely the frequency of development of chronic pain, acute stress reactions, satisfaction with the results of treatment and the number of wound localizations.MethodsThe treatment of 769 patients was analyzed. Pain intensity was diagnosed using a visual analog scale (VAS). To detect neuropathic pain, the Douleur Neuropathique 4 questions (DN4). The presence of an acute stress reaction (ASR) was diagnosed using The hospital anxiety and depression scale (HADS) and medical history, the diagnosis was established by a psychiatrist. Satisfaction with treatment results was studied using the Chaban quality of life scale (CQLS). Group comparisons were made using the Mann-Whitney test and the chi-square test, taking into account continuity correction.ResultsChronic pain was observed in 538 (70% 95% CI 66.7%-73.1%) patients with gunshot wounds: of them, 439 patients had wounds in 1, 2 anatomical parts of the body, here the frequency of pain chronicity is 69.7% (95% CI 66.0%-78.5%), and 99 patients had wounds in 3 or more anatomical parts of the body - 71.2% (95%CI 63.4%-78.5%). DN4 data suggest the presence of a neuropathic pain component in these patients. Also, all patients were diagnosed with ASR upon admission: the number of HADS points ranged from 9 to 25 points. CQLS data indicate that satisfaction with treatment outcomes was high (76 points) before hospital discharge, but subsequently decreased to a low level (64 points).ConclusionsPatients with gunshot wounds have a high risk of chronic pain, averaging 45% higher than the general population in civilian trauma patients. A greater frequency of the neuropathic component of pain and acute stress reactions is the reason for such chronicity. A decrease in the level of satisfaction with the results of treatment, in the remote period of observation, compared to the level at the time of discharge from the hospital, is probably a consequence of the formation of chronic pain.Trial registrationClinicalTrials.gov: Retrospectively registered on August 1, 2022, NCT05489029.

Dataset Information

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

Publications

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

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