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Article

Rapid Detection of Pathogens in Wound Exudate via Nucleic Acid Lateral Flow Immunoassay

1
Laboratory for MEMS Applications, IMTEK—Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany
2
Department of Trauma-, Hand- and Reconstructive Surgery, University Hospital Frankfurt, Johann Wolfgang Goethe University, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany
3
Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany
*
Author to whom correspondence should be addressed.
Biosensors 2021, 11(3), 74; https://doi.org/10.3390/bios11030074
Received: 4 February 2021 / Revised: 24 February 2021 / Accepted: 3 March 2021 / Published: 6 March 2021
(This article belongs to the Section Biosensors and Healthcare)
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. View Full-Text
Keywords: recombinase polymerase amplification; nucleic acid lateral flow immunoassay; point-of-care diagnostics; paper-based detection; nucleic acid amplification test; wound exudate; wound infection recombinase polymerase amplification; nucleic acid lateral flow immunoassay; point-of-care diagnostics; paper-based detection; nucleic acid amplification test; wound exudate; wound infection
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MDPI and ACS Style

Brunauer, A.; Verboket, R.D.; Kainz, D.M.; von Stetten, F.; Früh, S.M. Rapid Detection of Pathogens in Wound Exudate via Nucleic Acid Lateral Flow Immunoassay. Biosensors 2021, 11, 74. https://doi.org/10.3390/bios11030074

AMA Style

Brunauer A, Verboket RD, Kainz DM, von Stetten F, Früh SM. Rapid Detection of Pathogens in Wound Exudate via Nucleic Acid Lateral Flow Immunoassay. Biosensors. 2021; 11(3):74. https://doi.org/10.3390/bios11030074

Chicago/Turabian Style

Brunauer, Anna, René D. Verboket, Daniel M. Kainz, Felix von Stetten, and Susanna M. Früh 2021. "Rapid Detection of Pathogens in Wound Exudate via Nucleic Acid Lateral Flow Immunoassay" Biosensors 11, no. 3: 74. https://doi.org/10.3390/bios11030074

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