Development of CRISPR/Cas13a-based assays for the diagnosis of SchistosomiasisResearch in context
Skye R. MacGregor,
Donald P. McManus,
Haran Sivakumaran,
Thomas G. Egwang,
Moses Adriko,
Pengfei Cai,
Catherine A. Gordon,
Mary G. Duke,
Juliet D. French,
Natasha Collinson,
Remigio M. Olveda,
Gunter Hartel,
Carlos Graeff-Teixeira,
Malcolm K. Jones,
Hong You
Affiliations
Skye R. MacGregor
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Donald P. McManus
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Haran Sivakumaran
Genetics & Computational Biology Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Thomas G. Egwang
Department of Immunology and Parasitology, Med Biotech Laboratories, Kampala, Uganda
Moses Adriko
Vector Borne and NTD Control Division, Ministry of Health, Kampala, Uganda
Pengfei Cai
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Catherine A. Gordon
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia; School of Public Health, Faculty of Medicine, The University of Queensland, Brisbane, Australia
Mary G. Duke
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Juliet D. French
Genetics & Computational Biology Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Natasha Collinson
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
Remigio M. Olveda
Department of Health, Research Institute for Tropical Medicine, Manila, Philippines
Gunter Hartel
School of Public Health, Faculty of Medicine, The University of Queensland, Brisbane, Australia; Statistics, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia; School of Nursing, Queensland University of Technology, Brisbane, Queensland, Australia
Carlos Graeff-Teixeira
Department of Pathology, Infectious Diseases Unit, Health Sciences Center, Universidade Federal do Espírito Santo, Vitória, Brazil
Malcolm K. Jones
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia; School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia
Hong You
Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia; School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia; Corresponding author. Infection and Inflammation Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia.
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 National Health and Medical Research Council (NHMRC) of Australia (APP1194462, APP2008433).
