Affinity molecular assay for detecting Candida albicans using chitin affinity and RPA-CRISPR/Cas12a

  • Nat Commun. 2024 Oct 28;15(1):9304. doi: 10.1038/s41467-024-53693-5.
Shimei Shen  #  1  2  3  4  5 ,  Wen Wang  #  1  2  6 ,  Yuanyan Ma  #  1  4 ,  Shilei Wang  #  3 ,  Shaocheng Zhang  #  6  7 ,  Xuefei Cai  4 ,  Liang Chen  8 ,  Jin Zhang  1  4 ,  Yalan Li  1  4 ,  Xiaoli Wu  1  4 ,  Jie Wei  9 ,  Yanan Zhao  10 ,  Ailong Huang  11 ,  Siqiang Niu  12 ,  Deqiang Wang  13  14  15
Affiliations
  • 1. Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
  • 2. Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
  • 3. Department of Dermatology and Cosmetology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China.
  • 4. The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
  • 5. Department of Clinical Laboratory Medicine, Chongqing Red Cross Hospital (Jiangbei District People's Hospital), Chongqing, China.
  • 6. Department of Laboratory Medicine, The Second Affiliated Hospital of Chengdu Medical College (Nuclear Industry 416 Hospital), Chengdu, China.
  • 7. School of Clinical Medicine, Chengdu Medical College, Chengdu, China.
  • 8. Department of Pharmacy Practice, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, Buffalo, NY, USA.
  • 9. Department of Clinical Laboratory, Zhuhai People's Hospital (Zhuhai Hospital Affiliated with Jinan University), Zhuhai, China.
  • 10. Department of Pharmacy Practice, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, Buffalo, NY, USA. [email protected].
  • 11. The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China. [email protected].
  • 12. Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China. [email protected].
  • 13. Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China. [email protected].
  • 14. The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China. [email protected].
  • 15. Western (Chongqing) Collaborative Innovation Center for Intelligent Diagnostics and Digital Medicine, Chongqing National Biomedicine Industry Park, Chongqing, China. [email protected].
  • # Contributed equally.
PMID: 39468064 DOI: 10.1038/s41467-024-53693-5
Abstract

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.

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