dc.contributor.author | Cunningham, Clark H. | |
dc.contributor.author | Hennelly, Christopher M. | |
dc.contributor.author | Lin, Jessica T. | |
dc.contributor.author | Ubalee, Ratawan | |
dc.contributor.author | Boyc, Ross M. | |
dc.contributor.author | Mulogo, Edgar Mugema | |
dc.contributor.author | Hathaway, Nicholas | |
dc.contributor.author | Thwai, Kyaw L. | |
dc.contributor.author | Phanzu, Fernandine | |
dc.contributor.author | Kalonji, Albert | |
dc.contributor.author | Mwandagalirwa, Kashamuka | |
dc.contributor.author | Tshefu, Antoinette | |
dc.contributor.author | Juliano, Jonathan J. | |
dc.contributor.author | Parr, Jonathan B. | |
dc.date.accessioned | 2022-08-30T08:16:35Z | |
dc.date.available | 2022-08-30T08:16:35Z | |
dc.date.issued | 2021 | |
dc.identifier.citation | Cunningham, C. H., Hennelly, C. M., Lin, J. T., Ubalee, R., Boyce, R. M., Mulogo, E. M., ... & Parr, J. B. (2021). A novel CRISPR-based malaria diagnostic capable of Plasmodium detection, species differentiation, and drug-resistance genotyping. EBioMedicine, 68, 103415. | en_US |
dc.identifier.uri | http://ir.must.ac.ug/xmlui/handle/123456789/2424 | |
dc.description.abstract | Background: CRISPR-based diagnostics are a new class of highly sensitive and specific assays with multiple applications in infectious disease diagnosis. SHERLOCK, or Specific High-Sensitivity Enzymatic Reporter UnLOCKing, is one such CRISPR-based diagnostic that combines recombinase polymerase pre-amplification, CRISPR-RNA base-pairing, and LwCas13a activity for nucleic acid detection.
Methods: We developed SHERLOCK assays capable of detecting all Plasmodium species known to cause human malaria and species-specific detection of P. vivax and P. falciparum, the species responsible for the majority of malaria cases worldwide. We further tested these assays using a diverse panel of clinical samples from the Democratic Republic of the Congo, Uganda, and Thailand and pools of Anopheles mosquitoes from Thailand. In addition, we developed a prototype SHERLOCK assay capable of detecting the dihydropteroate synthetase (dhps) single nucleotide variant A581G associated with P. falciparum sulfadoxine resistance.
Findings: The suite of Plasmodium assays achieved analytical sensitivities ranging from 25-188 parasites per reaction when tested against laboratory strain genomic DNA. When compared to real-time PCR, the P. falciparum assay achieved 94% sensitivity and 94% specificity during testing of 123 clinical samples. Compared to amplicon-based deep sequencing, the dhps SHERLOCK assay achieved 73% sensitivity and 100% specificity when applied to a panel of 43 clinical samples, with false-negative calls only at lower parasite densities.
Interpretation: These novel SHERLOCK assays demonstrate the versatility of CRISPR-based diagnostics and their potential as a new generation of molecular tools for malaria diagnosis and surveillance | en_US |
dc.description.sponsorship | National Institutes of Health (T32GM007092, R21AI148579, K24AI134990, R01AI121558, UL1TR002489, P30CA016086) | en_US |
dc.language.iso | en_US | en_US |
dc.publisher | BioMedicine | en_US |
dc.subject | SHERLOCK | en_US |
dc.subject | Malaria | en_US |
dc.subject | CRISPR | en_US |
dc.subject | Cas13a | en_US |
dc.subject | Diagnostic | en_US |
dc.title | A novel CRISPR-based malaria diagnostic capable of Plasmodium detection, species differentiation, and drug-resistance genotyping | en_US |
dc.type | Article | en_US |