Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 May 19:6:34.
doi: 10.1038/s41421-020-0174-y. eCollection 2020.

SARS-CoV-2 detection with CRISPR diagnostics

Lu Guo  1   2   3 , Xuehan Sun #  1   2   3 , Xinge Wang #  1   2   3 , Chen Liang #  1   2   3 , Haiping Jiang #  1   2   3 , Qingqin Gao #  1   2   3 , Moyu Dai #  1   2   3 , Bin Qu  1   2   3 , Sen Fang  1   2   3 , Yihuan Mao  1   2   3 , Yangcan Chen  1   2   3 , Guihai Feng  1   2 , Qi Gu  1   2 , Ruiqi Rachel Wang  1   2 , Qi Zhou  1   2   3 , Wei Li  1   2   3
Affiliations

SARS-CoV-2 detection with CRISPR diagnostics

Lu Guo et al. Cell Discov. .
No abstract available

Keywords: Biological techniques; Molecular biology.

PubMed Disclaimer

Conflict of interest statement

Conflict of interestThe authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1. CASdetec used for SARS-CoV-2 detection.
a Fluorescence kinetics of sgRNA-3 for RdRp detection. E. coli cells bearing Blunt-SARS-CoV-RdRp or Blunt-SARS-CoV-2-RdRp were pre-incubated at 95 °C for 10 min and used as templates for RAA and CDetection. PAM sequences are colored in green, protospacers are colored in blue, base pair mismatches are colored in red. Error bars indicate standard errors of the mean (s.e.m.), n = 3. RFU, relative fluorescence units. b Fluorescence kinetics of RdRp detection using 108 nM sgRNA-3. Plasmid bearing SARS-CoV-2-RdRp was serially diluted as shown in the legend. n = 2. ΔRn, ΔFluorescence, which refers to the Rn value of an experimental reaction minus the Rn value of the baseline signal generated by ABI 7500. c Fluorescence kinetics of F1- and R1-based RdRp detection. SARS-CoV-2-RdRp RNA was serially diluted as shown in the legend. Error bars indicate (s.e.m.), n = 3. d Evaluation of cross-reactivity. Plamids containing target RdRp region from six human epidemic coronaviruses were serially diluted as the shown in the legend. n = 2. e Detection of SARS-CoV-2 pseudovirus. Virus genome was extracted using the virus RNA extraction kit (spin column). SARS-CoV were diluted to 5 × 105 copies/mL. n = 2. f Detection of SARS-CoV-2 pseudovirus. Virus was treated by direct lysis. SARS-CoV was diluted to 5 × 105 copies/mL. n = 2. g CASdetec results could be directly observed under blue LED. 3 replicates of products from Fig. 1c were imaged upon blue LED illumination. h Schematics showing the workflow of CASdetec. Virus genome was extracted by kit or direct lysis. Target sequences were pre-amplified by isothermal amplification, followed was CDetection. Fluorescence signals were obtained either from fluorescence reader or direct observation under blue light.
See this image and copyright information in PMC

References

    1. Coronaviridae Study Group of the International Committee on Taxonomy of Viruses. The species severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2. Nat. Microbiol. 2020;5:536–544. doi: 10.1038/s41564-020-0695-z. - DOI - PMC - PubMed
    1. Corman VM, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Eur. Surveill. 2020;25:2000045. doi: 10.2807/1560-7917.ES.2020.25.3.2000045. - DOI - PMC - PubMed
    1. Zhang, F., Abudayyeh, O. O. & Gootenberg, J. S. A protocol for detection of COVID-19 using CRISPR diagnostics. https://www.broadinstitute.org/files/publications/special/COVID-19%20det... (2020).
    1. Metsky, H. C. et al. CRISPR-based COVID-19 surveillance using a genomically-comprehensive machine learning approach. bioRxiv. 10.1101/2020.02.26.967026 (2020).
    1. Lucia, C., Federico, P. B. & Alejandra, G. C. An ultrasensitive, rapid, and portable coronavirus SARS-CoV-2 sequence detection method based on CRISPR-Cas12. bioRxiv. 10.1101/2020.02.29.971127 (2020).