Affinity molecular assay for detecting Candida albicans using chitin affinity and RPA-CRISPR/Cas12a
Invasive fungal infections (IFIs) pose a significant threat to immunocompromised individuals, leading to considerable morbidity and mortality. Prompt and accurate diagnosis is essential for effective treatment. Here we develop a rapid molecular diagnostic method that involves three steps: fungal enrichment using affinity-magnetic separation (AMS), genomic DNA extraction with silicon hydroxyl magnetic beads, and detection through a one-pot system. This method, optimized to detect 30 CFU/mL of C. albicans in blood and bronchoalveolar lavage (BAL) samples within 2.5 h, is approximately 100 times more sensitive than microscopy-based staining. Initial validation using clinical samples showed 93.93% sensitivity, 100% specificity, and high predictive values, while simulated tests demonstrated 95% sensitivity and 100% specificity. This cost-effective, highly sensitive technique offers potential for use in resource-limited clinical settings and can be easily adapted to differentiate between fungal species and detect drug resistance.
Authors
- Shilei Wang (ORCID: https://orcid.org/0000-0002-3343-9867)
- Ailong Huang (ORCID: https://orcid.org/0000-0003-0148-7423)
- Yuanyan Ma
- Deqiang Wang
- Siqiang Niu (ORCID: https://orcid.org/0000-0002-8911-7138)
- Jie Wei (ORCID: https://orcid.org/0009-0002-2403-3889)
- Yanan Zhao (ORCID: https://orcid.org/0000-0002-6634-2949)
- Liang Chen (ORCID: https://orcid.org/0000-0001-5845-2235)
- Yalan Li (ORCID: https://orcid.org/0009-0006-0300-9723)
- Xiaoli Wu (ORCID: https://orcid.org/0009-0008-5078-2158)
- Wen Wang (ORCID: https://orcid.org/0000-0002-5812-6744)
- Shaocheng Zhang (ORCID: https://orcid.org/0000-0003-3620-1616)
- Shimei Shen
- Xue-Fei Cai
- Jin Zhang (ORCID: https://orcid.org/0000-0003-3554-2996)
Institutions
- Jinan University (CN)
- Dalian Medical University (CN)
- Chengdu Medical College (CN)
- Runze (China) (CN)
- Ministry of Education (SA)
- Second Affiliated Hospital of Chongqing Medical University (CN)
- Zhuhai People's Hospital (CN)
- Ministry of Education (TH)
- The Affiliated Yongchuan Hospital of Chongqing Medical University (CN)
- Children's Hospital of Chongqing Medical University (CN)
- First People's Hospital of Chongqing (CN)
- University at Buffalo, State University of New York (US)
- Chongqing Medical University (CN)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-10-28
- DOI
- https://doi.org/10.1038/s41467-024-53693-5
- Citations
- 31
- Primary Topic
- Biosensors and Analytical Detection
- Type
- article
- Field-Weighted Citation Impact
- 5.72
Funders
- National Science Foundation
- National Natural Science Foundation of China
- Chongqing Medical University
- Natural Science Foundation of Chongqing
- Centre Scientifique et Technique du Bâtiment
- National Science and Technology Major Project
- Natural Science Foundation of Sichuan Province
- Basic and Applied Basic Research Foundation of Guangdong Province