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Digital CRISPR/Cas-Assisted Assay for Rapid and Sensitive Detection of SARS-CoV-2. | LitMetric

AI Article Synopsis

  • The COVID-19 pandemic has highlighted the need for rapid diagnostic tests, particularly those utilizing CRISPR/Cas systems for nucleic acid detection.
  • A new assay called deCOViD was developed, integrating digital technology with CRISPR/Cas12a-assisted methods, enabling quick and sensitive detection of SARS-CoV-2 within 15-30 minutes.
  • deCOViD offers significant advantages, including high sensitivity to low concentrations of viral RNA and the ability to analyze clinical samples, making it one of the fastest and most effective CRISPR-based detection methods available.

Article Abstract

The unprecedented demand for rapid diagnostics in response to the COVID-19 pandemic has brought the spotlight onto clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated systems (Cas)-assisted nucleic acid detection assays. Already benefitting from an elegant detection mechanism, fast assay time, and low reaction temperature, these assays can be further advanced via integration with powerful, digital-based detection. Thus motivated, the first digital CRISPR/Cas-assisted assay-coined digitization-enhanced CRISPR/Cas-assisted one-pot virus detection (deCOViD)-is developed and applied toward SARS-CoV-2 detection. deCOViD is realized through tuning and discretizing a one-step, fluorescence-based, CRISPR/Cas12a-assisted reverse transcription recombinase polymerase amplification assay into sub-nanoliter reaction wells within commercially available microfluidic digital chips. The uniformly elevated digital concentrations enable deCOViD to achieve qualitative detection in <15 min and quantitative detection in 30 min with high signal-to-background ratio, broad dynamic range, and high sensitivity-down to 1 genome equivalent (GE) µL of SARS-CoV-2 RNA and 20 GE µL of heat-inactivated SARS-CoV-2, which outstrips its benchtop-based counterpart and represents one of the fastest and most sensitive CRISPR/Cas-assisted SARS-CoV-2 detection to date. Moreover, deCOViD can detect RNA extracts from clinical samples. Taken together, deCOViD opens a new avenue for advancing CRISPR/Cas-assisted assays and combating the COVID-19 pandemic and beyond.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7927608PMC
http://dx.doi.org/10.1002/advs.202003564DOI Listing

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