Field-deployable Viral Diagnostics Using CRISPR-Cas13

Science. 2018 Apr 27;360(6387):444-448. doi: 10.1126/science.aas8836.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological / genetics
  • Bacterial Proteins / chemistry*
  • CRISPR-Associated Proteins / chemistry*
  • Dengue / diagnosis*
  • Dengue Virus / genetics
  • Dengue Virus / isolation & purification*
  • Endonucleases / chemistry*
  • Enzyme Assays*
  • Humans
  • Microcephaly / diagnosis
  • Microcephaly / virology
  • Polymorphism, Single Nucleotide
  • RNA, Viral / analysis*
  • Zika Virus / genetics
  • Zika Virus / isolation & purification*
  • Zika Virus Infection / diagnosis*

Substances

  • Bacterial Proteins
  • CRISPR-Associated Proteins
  • RNA, Viral
  • Endonucleases