Summary Background Schistosomiasis is a disease that significantly impacts human health in the developing world. Effective diagnostics are urgently needed for improved control of this disease. CRISPR-based technology has rapidly accelerated the development of a revolutionary and powerful diagnostics platform, resulting in the advancement of a class of ultrasensitive, specific, cost-effective and portable diagnostics, typified by applications in COVID-19/cancer diagnosis. Methods We developed CRISPR-based diagnostic platform SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) for the detection of Schistosoma japonicum and S. mansoni by combining recombinase polymerase amplification (RPA) with CRISPR-Cas13a detection, measured via fluorescent or colorimetric readouts. We evaluated SHERLOCK assays by using 150 faecal/serum samples collected from Schistosoma -infected ARC Swiss mice (female), and 189 human faecal/serum samples obtained from a S. japonicum- endemic area in the Philippines and a S. mansoni- endemic area in Uganda. Findings The S. japonicum SHERLOCK assay achieved 93–100% concordance with gold-standard qPCR detection across all the samples. The S. mansoni SHERLOCK assay demonstrated higher sensitivity than qPCR and was able to detect infection in mouse serum as early as 3 weeks post-infection. In human samples, S. mansoni SHERLOCK had 100% sensitivity when compared to qPCR of faecal and serum samples. Interpretation These schistosomiasis diagnostic assays demonstrate the potential of SHERLOCK/CRISPR-based diagnostics to provide highly accurate and field-friendly point-of-care tests that could provide the next generation of diagnostic and surveillance tools for parasitic neglected tropical diseases. Funding Australian Infectious Diseases Research Centre seed grant (2022) and 10.13039/501100000925 National Health and Medical Research Council (NHMRC) of Australia (APP1194462, APP2008433).
【저자키워드】 diagnostics, SHERLOCK, CRISPR, point-of-care, Schistosomiasis, Schistosoma mansoni, Schistosoma japonicum, Cas13a,