CRISPR based assay

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Original Article
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A RT loop mediated isothermal amplification technology combined with CRISPR-Cas12 based detection of SARS-Cov-2 showed improved specificity against N and E genes and assay sensitivity. Additional information: (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7724759/bin/ac0c04047_si_001.pdf)
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 		33238709
(Anal Chem)
PMID
33238709
Date of Publishing: 2020 Dec 15
Title Isothermal Amplification and Ambient Visualization in a Single Tube for the Detection of SARS-CoV-2 Using Loop-Mediated Amplification and CRISPR technology
Author(s) namePang B, Xu J et al.
Journal Anal Chem
Impact factor
6.37
Citation count: 41

Reporting SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) method to detect SARS-CoV-2, which relies on the RT-RPA method to amplify viral gene segments, followed by CRISPR-Cas13a-mediated detection of amplified genes. The assay was 100% specific, and 96% sensitive with the fluorescence readout, and 88% sensitive with the lateral-flow readout. 1. SHERLOCK-based detection of the SARS-CoV-2 S gene is specific to SARS-CoV-2, with no cross-reactivity towards other common human coronaviruses 2. Table S3. RPA primers, crRNAs, and RNA reporters information(https://static-content.springer.com/esm/art%3A10.1038%2Fs41551-020-00603-x/MediaObjects/41551_2020_603_MOESM1_ESM.pdf)
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 		32848209
(Nat Biomed Eng)
PMID
32848209
Date of Publishing: 2020 Dec
Title Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA
Author(s) namePatchsung M, Jantarug K et al.
Journal Nat Biomed Eng
Impact factor
31.8
Citation count: 138

Reporting a simple STOP (SHERLOCK testing in one pot assay) diagnostic method for the detection of the SARS-CoV-2 N gene. This method combines viral RNA extraction with isothermal amplification and CRISPR-mediated detection technology. The STOPCovid.v2 test has a detection limit of 40.3 and a false negative test Ct value greater than 37. Also the detection time for the result is about 15-45 minutes.
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 		32937062
(N Engl J Med)
PMID
32937062
Date of Publishing: 2020 Oct 8
Title Detection of SARS-CoV-2 with SHERLOCK One-Pot Testing
Author(s) nameJoung J, Ladha A et al.
Journal N Engl J Med
Impact factor
37.91
Citation count: 162

In this study, iSCAN assay (in vitro Specific CRISPR-based Assay for Nucleic acids) is developed, involving RT-LAMP coupled with CRISPR-Cas12, for rapid, accurate, specific, sensitive detection of COVID-19. Detailed information about the assay protocol, primer and result is provided in Supplementary data (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7434412/bin/mmc1.docx)
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 		32822689
(Virus Res)
PMID
32822689
Date of Publishing: 2020 Oct 15
Title iSCAN: An RT-LAMP-coupled CRISPR-Cas12 module
Author(s) nameAli Z, Aman R et al.
Journal Virus Res
Impact factor
2.6
Citation count: 81

Reporting a one-step RT-AIOD-CRISPR assay based on a unique dual CRISPR-Cas12a detection method for the rapid detection of SARS-COV-2. The AIOD-CRISPR assay is lost cost assay (~$6), a color change from orange-yellow to green indicates the positive result.
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 		32948757
(Nat Commun)
PMID
32948757
Date of Publishing: 2020 Sep 18
Title Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay
Author(s) nameDing X, Yin K et al.
Journal Nat Commun
Impact factor
11.8
Citation count: 128

In this study, a rapid, accurate CRISPR-Cas12-based lateral flow assay is developed to detect SARS-CoV-2 from clinical samples. The CRISPR-based DETECTR technology provides a faster alternative to the US CDC RT-qPCR assay, with 95% positive predictive agreement and 100% negative predictive agreement. 1. Tests such as the DETECTR assay reported here are amenable to periodic repeat testing of patient samples 2. We report that our CRISPR-based DETECTR technology can be reconfigured within days to detect SARS-CoV-2
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 		32300245
(Nat Biotechnol)
PMID
32300245
Date of Publishing: 2020 Jul
Title CRISPR-Cas12-based detection of SARS-CoV-2
Author(s) nameBroughton JP, Deng X et al.
Journal Nat Biotechnol
Impact factor
36.9
Citation count: 684

In this study, a CRISPR-assisted detection protocol (CASdetec) is developed and optimized for SARS-CoV-2 detection. The protocol includes virus handling, nucleic acid amplification by RT-RAA, and CRISPR-based detection with a blue LED for positive samples. This assay can detect the virus with at least 1 10^4 copies/mL, with no cross-reactivity with the other human coronaviruses. Primer details are provided in Supplementary Table S1 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7235268/#MOESM1).
Out of 1792 sequences analyzed from GISAID, 1673 sequences were matching to chosen primers and sgRNA, suggesting that selected sgRNA and primers can be used for nearly all of the reported SARS-CoV-2 genomes.
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 		32435508
(Cell Discov)
PMID
32435508
Date of Publishing: 2020
Title SARS-CoV-2 detection with CRISPR diagnostics
Author(s) nameGuo L, Sun X et al.
Journal Cell Discov
Impact factor
4.65
Citation count: 70