ORIGINAL RESEARCH article
Front. Chem.
Sec. Analytical Chemistry
Volume 13 - 2025 | doi: 10.3389/fchem.2025.1654270
This article is part of the Research TopicChemistry and Materials in Lateral Flow AssayView all articles
Rapid detection of Pseudomonas aeruginosa by glycerol one-pot RAA/CRISPR-Cas12a method
Provisionally accepted
- 1Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, China
- 2Baise People's Hospital; Affiliated Southwest Hospital of Youjiang Medical University for Nationalities, Baise, China
- 3Yaneng BlOscience (Shenzhen) Corporation, Guangdong, China
- 4Clinical Genome Center, Guangxi KingMed Diagnostics, Guangxi, China
- 5Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
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Pseudomonas aeruginosa (PA), an opportunistic pathogen commonly responsible for hospital-acquired infections, poses significant threats to human health.To enable rapid and reliable PA detection while effectively mitigating aerosol contamination risks inherent in conventional methods. We developed a glycerol one-pot Recombinase-aided Amplification (RAA)/CRISPR-Cas12a method. Four result reading methods were established: Fluorescence Detection (FD), Blue Light Irradiation Detection (BLD), and Ultraviolet Irradiation Detection (UID), as well as Lateral Flow Chromatography Strip (LFS). The glycerol one-pot RAA-CRISPR/Cas12a method demonstrated high specificity and sensitivity in detecting the PA-specific lasB gene. The detection limit reached 1.20 × 10⁻⁴ng/μL (fluorescence-based) and 1.20 × 10⁻³ ng/μL (LFS-based). In validation against 64 clinical isolates, compared to conventional PCR, the assay achieved 100% sensitivity, specificity, and accuracy in lasB detection. In conclusion, the glycerol one-pot RAA/CRISPR-Cas12a method provides a rapid, sensitive, and straightforward platform, providing a promising approach for clinical diagnosis of PA and environmental surveillance applications.
Keywords: Recombinase-aided amplification, glycerol one-pot CRISPR/Cas12a method, lateral flow chromatography strip, Rapid detection, Pseudomonas aeruginosa
Received: 26 Jun 2025; Accepted: 15 Jul 2025.
Copyright: © 2025 Wei, Luo, Zhou, Ren, Li, Liang, Li and Wei. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Lina Liang, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, China
Xuebin Li, Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
Guijiang Wei, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, China
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