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Field-deployable viral diagnostics using CRISPR-Cas13
Cameron Myhrvold1,2, Catherine A Freije1,2,3, Jonathan S Gootenberg4,5,6,7,8
1Broad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA. pardis@broadinstitute.org cmyhrvol@broadinstitute.org cfreije@broadinstitute.org.
Science (New York, N.Y.)
|April 28, 2018
Summary
A new diagnostic platform, SHERLOCK (specific high-sensitivity enzymatic reporter unlocking), rapidly detects Zika virus (ZIKV) and dengue virus (DENV) in patient samples. The HUDSON (heating unextracted diagnostic samples to obliterate nucleases) protocol enables instrument-free detection in under two hours.
Area of Science:
- Molecular Biology
- Virology
- Biotechnology
Background:
- Global infectious disease mitigation necessitates sensitive, specific, and field-deployable diagnostic tools.
- Existing diagnostic methods may lack the speed and accessibility required for rapid outbreak response.
Purpose of the Study:
- To evaluate the Cas13-based SHERLOCK platform for sensitive and specific detection of Zika virus (ZIKV) and dengue virus (DENV).
- To develop and validate the HUDSON protocol for rapid, instrument-free viral detection directly from patient samples.
- To assess SHERLOCK's capability in differentiating DENV serotypes and ZIKV strains.
Main Methods:
- Utilized the Cas13-based SHERLOCK platform for nucleic acid detection.
- Developed and implemented the HUDSON protocol for sample preparation.
- Tested assay sensitivity down to 1 copy per microliter for ZIKV and DENV.
- Validated SHERLOCK's ability to distinguish DENV serotypes and ZIKV strains.
Main Results:
- SHERLOCK detected ZIKV and DENV at concentrations as low as 1 copy/µL.
- The HUDSON protocol enabled instrument-free DENV detection from patient samples in under 2 hours.
- SHERLOCK successfully differentiated the four DENV serotypes and pandemic ZIKV strains.
- Rapid (<1 week) design and testing of instrument-free assays for viral single-nucleotide polymorphisms were achieved.
Conclusions:
- The SHERLOCK platform, combined with the HUDSON protocol, offers a sensitive, specific, and rapid diagnostic solution for ZIKV and DENV.
- This approach facilitates instrument-free, point-of-care detection, crucial for managing infectious disease outbreaks.
- The platform's ability to detect specific viral variants and single-nucleotide polymorphisms enhances its utility in molecular epidemiology.
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