文章基本信息

标题：Rapid Detection of Pathogens in Wound Exudate via Nucleic Acid Lateral Flow Immunoassay
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作者：Anna Brunauer ; René D. Verboket ; Daniel M. Kainz 等
期刊名称：Biosensors
电子版ISSN：2079-6374
出版年度：2021
卷号：11
期号：3
页码：74
DOI：10.3390/bios11030074
出版社：MDPI Publishing
摘要：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.
关键词：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