@article{Wang:2022:10.1016/j.snb.2022.131765,
author = {Wang, Y and Xue, T and Wang, M and Ledesma-Amaro, R and Lu, Y and Hu, X and Zhang, T and Yang, M and Li, Y and Xiang, J and Deng, R and Ying, B and Li, W},
doi = {10.1016/j.snb.2022.131765},
journal = {Sensors and Actuators B: Chemical: international journal devoted to research and development of physical and chemical transducers},
title = {CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples},
url = {http://dx.doi.org/10.1016/j.snb.2022.131765},
volume = {362},
year = {2022}
}

Download

TY  - JOUR
AB  - SARS-CoV-2 is one of the greatest threats to global human health. Point-of-care diagnostic tools for SARS-CoV-2 could facilitate rapid therapeutic intervention and mitigate transmission. In this work, we report CRISPR-Cas13a cascade-based viral RNA (Cas13C) assay for label-free and isothermal determination of SARS-CoV-2 and its mutations in clinical samples. Cas13a/crRNA was utilized to directly recognize the target of SARS-CoV-2 RNA, and the recognition events sequentially initiate the transcription amplification to produce light-up RNA aptamers for output fluorescence signal. The recognition of viral RNA via Cas13a-guide RNA ensures a high specificity to distinguish SARS-CoV-2 from MERS-CoV and SARS-CoV, as well as viral mutations. A post transcription amplification strategy was triggered after CRISPR-Cas13a recognition contributes to an amplification cascade that achieves high sensitivity for detecting SARS-CoV-2 RNA, with a limit of detection of 0.216 fM. In addition, the Cas13C assay could be able to discriminate single-nucleotide mutation, which was proven with N501Y in SARS-Cov-2 variant. This method was validated by a 100% agreement with RT-qPCR results from 12 clinical throat swab specimens. The Cas13C assay has the potential to be used as a routine nucleic acid test of SARS-CoV-2 virus in resource-limited regions.
AU  - Wang,Y
AU  - Xue,T
AU  - Wang,M
AU  - Ledesma-Amaro,R
AU  - Lu,Y
AU  - Hu,X
AU  - Zhang,T
AU  - Yang,M
AU  - Li,Y
AU  - Xiang,J
AU  - Deng,R
AU  - Ying,B
AU  - Li,W
DO  - 10.1016/j.snb.2022.131765
PY  - 2022///
SN  - 0925-4005
TI  - CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples
T2  - Sensors and Actuators B: Chemical: international journal devoted to research and development of physical and chemical transducers
UR  - http://dx.doi.org/10.1016/j.snb.2022.131765
UR  - https://www.ncbi.nlm.nih.gov/pubmed/35370361
UR  - http://hdl.handle.net/10044/1/96215
VL  - 362
ER  - 

Download