Field-deployable viral diagnostics using CRISPR-Cas13
By C. Myhrvold, C. A. Freije, Jonathan Gootenberg, Omar Abudayyeh, Hayden C. Metsky, A. Durbin, M. Kellner, A. L. Tan, Lauren M. Paul, L. Parham and 14 more. Science, 2018
1,250 citations recorded by the source corpus.
Taking CRISPR technology further CRISPR techniques are allowing the development of technologies for nucleic acid detection (see the Perspective by Chertow). Taking advantages of the distinctive enzymatic properties of CRISPR enzymes, Gootenberg et al. developed an improved nucleic acid detection technology for multiplexed quantitative and highly sensitive detection, combined with lateral flow for visual readout. Myhrvold et al. added a sample preparation protocol to create a field-deployable viral diagnostic platform for rapid detection of specific strains of pathogens in clinical samples. Cas12a (also known as Cpf1), a type V CRISPR protein, cleaves double-stranded DNA and has been adapted for genome editing. Chen et al. discovered that Cas12a also processes single-stranded DNA threading activity. A technology platform based on this activity detected human papillomavirus in patient samples with high sensitivity. Science, this issue p. 439, p. 444, p. 436; see also p. 381 A nucleic acid detection technology identifies viruses with minimal equipment and sample processing requirements. Mitigating global infectious disease requires diagnostic tools that are sensitive, specific, and rapidly field deployable. In this study, we demonstrate that the Cas13-based SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform can detect Zika virus (ZIKV) and dengue virus (DENV) in patient samples at concentrations as low as 1 copy per microliter. We developed HUDSON (heating unextracted diagnostic samples to obliterate nucleases), a protocol that pairs with SHERLOCK for viral detection directly from bodily fluids, enabling instrument-free DENV detection directly from patient samples in <2 hours. We further demonstrate that SHERLOCK can distinguish the four DENV serotypes, as well as region-specific strains of ZIKV from the 2015–2016 pandemic. Finally, we report the rapid (<1 week) design and testing of instrument-free assays to detect clinically relevant viral single-nucleotide polymorphisms.
Subfields: Epidemiology, Molecular Biology, Public Health, Environmental and Occupational Health
View Paper: https://www.semanticscholar.org/paper/6b149fdae50eb9797b6f4757061d8e37faea1a48
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Citations in the corpus
- Virology, Epidemiology, Pathogenesis, and Control of COVID-19 (2020, methodology): “The Cas13-based SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform has been widely used to detect Zika virus (ZIKV) and dengue virus (DENV) in patient samples at concentrations as low as 1 copy per microliter [47].”
- CRISPR/Cas Systems towards Next-Generation Biosensing. (2019, methodology): “Cas12b with LAMP amplifica on [40] Cas12a with RPA amplifica on [36] Update with raw sample dectec on capaci es [19]”
- Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy (2020, neutral): “, 2019) is now being harnessed for viral diagnostics (Chen et al., 2018; East-Seletsky et al., 2016; Gootenberg et al., 2018; 2017; Myhrvold et al., 2018).”