Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay

Nat Commun. 2020 Sep 18;11(1):4711. doi: 10.1038/s41467-020-18575-6.


The recent outbreak of novel coronavirus (SARS-CoV-2) causing COVID-19 disease spreads rapidly in the world. Rapid and early detection of SARS-CoV-2 facilitates early intervention and prevents the disease spread. Here, we present an All-In-One Dual CRISPR-Cas12a (AIOD-CRISPR) assay for one-pot, ultrasensitive, and visual SARS-CoV-2 detection. By targeting SARS-CoV-2's nucleoprotein gene, two CRISPR RNAs without protospacer adjacent motif (PAM) site limitation are introduced to develop the AIOD-CRISPR assay and detect the nucleic acids with a sensitivity of few copies. We validate the assay by using COVID-19 clinical swab samples and obtain consistent results with RT-PCR assay. Furthermore, a low-cost hand warmer (~$0.3) is used as an incubator of the AIOD-CRISPR assay to detect clinical samples within 20 min, enabling an instrument-free, visual SARS-CoV-2 detection at the point of care. Thus, our method has the significant potential to provide a rapid, sensitive, one-pot point-of-care assay for SARS-CoV-2.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Betacoronavirus / genetics
  • Betacoronavirus / isolation & purification*
  • COVID-19
  • COVID-19 Testing
  • CRISPR-Cas Systems
  • Clinical Laboratory Techniques / methods
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / virology*
  • Genes, Viral
  • Humans
  • Molecular Diagnostic Techniques / methods
  • Nucleic Acid Amplification Techniques / methods
  • Pandemics
  • Pneumonia, Viral / diagnosis
  • Pneumonia, Viral / virology*
  • Point-of-Care Systems
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • SARS-CoV-2
  • Sensitivity and Specificity
  • Viral Proteins / analysis
  • Viral Proteins / genetics


  • RNA, Viral
  • Viral Proteins