Frontiers in Chemistry (Jul 2025)
Rapid detection of Pseudomonas aeruginosa by glycerol one-pot RAA/CRISPR-Cas12a method
- Lijian Wei,
- Lijian Wei,
- Lijian Wei,
- Lijian Wei,
- Lijian Wei,
- Lijian Wei,
- Shihua Luo,
- Shihua Luo,
- Shihua Luo,
- Shihua Luo,
- Shihua Luo,
- Weijie Zhou,
- Baoyan Ren,
- Baoyan Ren,
- Miao Li,
- Lina Liang,
- Lina Liang,
- Lina Liang,
- Lina Liang,
- Lina Liang,
- Xuebin Li,
- Xuebin Li,
- Xuebin Li,
- Xuebin Li,
- Guijiang Wei,
- Guijiang Wei,
- Guijiang Wei,
- Guijiang Wei,
- Guijiang Wei,
- Guijiang Wei
Affiliations
- Lijian Wei
- Center for Medical Laboratory Science, Affiliated Hospital of Youjiang Medical University for Nationalities, Guangxi, China
- Lijian Wei
- Key Laboratory of Research on Clinical Molecular Diagnosis for High Incidence Diseases in Western Guangxi of Guangxi Higher Education Institutions, Guangxi, China
- Lijian Wei
- Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Lijian Wei
- Engineering Research Center of Guangxi Higher Education Institutions for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Lijian Wei
- Guangxi Engineering Research Center for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Lijian Wei
- Department of Clinical Laboratory, Baise People’s Hospital, Affiliated Southwest Hospital of Youjiang Medical University for Nationalities, Guangxi, China
- Shihua Luo
- Center for Medical Laboratory Science, Affiliated Hospital of Youjiang Medical University for Nationalities, Guangxi, China
- Shihua Luo
- Key Laboratory of Research on Clinical Molecular Diagnosis for High Incidence Diseases in Western Guangxi of Guangxi Higher Education Institutions, Guangxi, China
- Shihua Luo
- Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Shihua Luo
- Engineering Research Center of Guangxi Higher Education Institutions for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Shihua Luo
- Guangxi Engineering Research Center for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Weijie Zhou
- Department of Clinical Laboratory, Baise People’s Hospital, Affiliated Southwest Hospital of Youjiang Medical University for Nationalities, Guangxi, China
- Baoyan Ren
- Yaneng BlOscience (Shenzhen) Corporation, Guangdong, China
- Baoyan Ren
- Modern Industrial College of Biomedicine and Great Health, Youjiang Medical University for Nationalities, Guangxi, China
- Miao Li
- Clinical Genome Center, Guangxi KingMed Diagnostics, Guangxi, China
- Lina Liang
- Center for Medical Laboratory Science, Affiliated Hospital of Youjiang Medical University for Nationalities, Guangxi, China
- Lina Liang
- Key Laboratory of Research on Clinical Molecular Diagnosis for High Incidence Diseases in Western Guangxi of Guangxi Higher Education Institutions, Guangxi, China
- Lina Liang
- Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Lina Liang
- Engineering Research Center of Guangxi Higher Education Institutions for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Lina Liang
- Guangxi Engineering Research Center for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Xuebin Li
- Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Xuebin Li
- Engineering Research Center of Guangxi Higher Education Institutions for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Xuebin Li
- Guangxi Engineering Research Center for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Xuebin Li
- Modern Industrial College of Biomedicine and Great Health, Youjiang Medical University for Nationalities, Guangxi, China
- Guijiang Wei
- Center for Medical Laboratory Science, Affiliated Hospital of Youjiang Medical University for Nationalities, Guangxi, China
- Guijiang Wei
- Key Laboratory of Research on Clinical Molecular Diagnosis for High Incidence Diseases in Western Guangxi of Guangxi Higher Education Institutions, Guangxi, China
- Guijiang Wei
- Baise Key Laboratory for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Guijiang Wei
- Engineering Research Center of Guangxi Higher Education Institutions for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Guijiang Wei
- Guangxi Engineering Research Center for Precise Genetic Testing of Long-dwelling Nationalities, Guangxi, China
- Guijiang Wei
- Modern Industrial College of Biomedicine and Great Health, Youjiang Medical University for Nationalities, Guangxi, China
- DOI
- https://doi.org/10.3389/fchem.2025.1654270
- Journal volume & issue
-
Vol. 13
Abstract
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 × 10-4 ng/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
