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

• Published in: Nature communications 2024-10, Vol.15 (1), p.9304-16, Article 9304
• Main Authors: Shen, Shimei, Wang, Wen, Ma, Yuanyan, Wang, Shilei, Zhang, Shaocheng, Cai, Xuefei, Chen, Liang, Zhang, Jin, Li, Yalan, Wu, Xiaoli, Wei, Jie, Zhao, Yanan, Huang, Ailong, Niu, Siqiang, Wang, Deqiang
• Format: Article
• Language: English
• Subjects: 59; 631/208/4041; 631/61/32; 692/53/2421; 692/699/255/1672; Affinity; Bacterial Proteins; Bronchoalveolar Lavage Fluid - microbiology; Bronchus; Candida albicans - genetics; Candida albicans - isolation & purification; Candidiasis - diagnosis; Candidiasis - microbiology; Chitin; Chitin - metabolism; CRISPR; CRISPR-Associated Proteins; CRISPR-Cas Systems; Diagnostic systems; DNA, Fungal - genetics; Drug resistance; Endodeoxyribonucleases - genetics; Endodeoxyribonucleases - metabolism; Fungi; Humanities and Social Sciences; Humans; Lavage; Magnetic separation; Morbidity; multidisciplinary; Science; Science (multidisciplinary); Sensitivity; Sensitivity and Specificity
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-53693-5

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. IFIs pose a significant threat to immunocompromised individuals, and there is an urgent need for rapid and sensitive diagnostic assays. Here, the authors present a rapid molecular diagnostic method based on magnetic fungal enrichment and CRISPR-based detection.