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Genome-wide tiled detection of circulating Mycobacterium tuberculosis cell-free DNA using Cas13. Nature communications Thakku, S. G., Lirette, J., Murugesan, K., Chen, J., Theron, G., Banaei, N., Blainey, P. C., Gomez, J., Wong, S. Y., Hung, D. T. 2023; 14 (1): 1803

Abstract

Detection of microbial cell-free DNA (cfDNA) circulating in the bloodstream has emerged as a promising new approach for diagnosing infection. Microbial diagnostics based on cfDNA require assays that can detect rare and highly fragmented pathogen nucleic acids. We now report WATSON (Whole-genome Assay using Tiled Surveillance Of Nucleic acids), a method to detect low amounts of pathogen cfDNA that couples pooled amplification of genomic targets tiled across the genome with pooled CRISPR/Cas13-based detection of these targets. We demonstrate that this strategy of tiling improves cfDNA detection compared to amplification and detection of a single targeted locus. WATSON can detect cfDNA from Mycobacterium tuberculosis in plasma of patients with active pulmonary tuberculosis, a disease that urgently needs accurate, minimally-invasive, field-deployable diagnostics. We thus demonstrate the potential for translating WATSON to a lateral flow platform. WATSON demonstrates the ability to capitalize on the strengths of targeting microbial cfDNA to address the need for point-of-care diagnostic tests for infectious diseases.

View details for DOI 10.1038/s41467-023-37183-8

View details for PubMedID 37002219

View details for PubMedCentralID PMC10064635