Rapid detection of pathogens in wound exudate via nucleic acid lateral flow immunoassay

  • The rapid detection of pathogens in infected wounds can significantly improve the clinical outcome. Wound exudate, which can be collected in a non-invasive way, offers an attractive sample material for the detection of pathogens at the point-of-care (POC). Here, we report the development of a nucleic acid lateral flow immunoassay for direct detection of isothermally amplified DNA combined with fast sample preparation. The streamlined protocol was evaluated using human wound exudate spiked with the opportunistic pathogen Pseudomonas aeruginosa that cause severe health issues upon wound colonization. A detection limit of 2.1 × 105 CFU per mL of wound fluid was achieved, and no cross-reaction with other pathogens was observed. Furthermore, we integrated an internal amplification control that excludes false negative results and, in combination with the flow control, ensures the validity of the test result. The paper-based approach with only three simple hands-on steps has a turn-around time of less than 30 min and covers the complete analytical process chain from sample to answer. This newly developed workflow for wound fluid diagnostics has tremendous potential for reliable pathogen POC testing and subsequent target-oriented therapy.

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Author:Anna Brunauer, René D. VerboketORCiDGND, Daniel M. Kainz, Felix von Stetten, Susanna Maria Früh
Parent Title (English):Biosensors
Place of publication:Basel
Document Type:Article
Date of Publication (online):2021/03/06
Date of first Publication:2021/03/06
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2021/10/19
Tag:nucleic acid amplification test; nucleic acid lateral flow immunoassay; paper-based detection; point-of-care diagnostics; recombinase polymerase amplification; wound exudate; wound infection
Issue:3, art. 74
Page Number:16
First Page:1
Last Page:16
Funding from the German Research Foundation (DFG), within the project “Hochdynamis- che Multianalyte-Assays” (grant number 397660978), is gratefully acknowledged.
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung 4.0