Back to Search
Start Over
Detection of Infectious Viruses Using CRISPR-Cas12-Based Assay.
- Source :
-
Biosensors [Biosensors (Basel)] 2021 Aug 28; Vol. 11 (9). Date of Electronic Publication: 2021 Aug 28. - Publication Year :
- 2021
-
Abstract
- The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease-19 (COVID-19), has severely influenced public health and economics. For the detection of SARS-CoV-2, clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein (Cas)-based assays have been emerged because of their simplicity, sensitivity, specificity, and wide applicability. Herein, we have developed a CRISPR-Cas12-based assay for the detection of SARS-CoV-2. In the assay, the target amplicons are produced by isothermal reverse transcription recombinase polymerase amplification (RT-RPA) and recognized by a CRISPR-Cas12a/guide RNA (gRNA) complex that is coupled with the collateral cleavage activity of fluorophore-tagged probes, allowing either a fluorescent measurement or naked-eye detection on a lateral flow paper strip. This assay enables the sensitive detection of SARS-CoV-2 at a low concentration of 10 copies per sample. Moreover, the reliability of the method is verified by using nasal swabs and sputum of COVID-19 patients. We also proved that the current assay can be applied to other viruses, such as Middle East respiratory syndrome coronavirus (MERS-CoV) and severe acute respiratory syndrome coronavirus (SARS-CoV), with no major changes to the basic scheme of testing. It is anticipated that the CRISPR-Cas12-based assay has the potential to serve as a point-of-care testing (POCT) tool for a wide range of infectious viruses.
- Subjects :
- CRISPR-Cas Systems
Fluorescent Dyes chemistry
Humans
Middle East Respiratory Syndrome Coronavirus genetics
Nose virology
Point-of-Care Testing
RNA, Guide, CRISPR-Cas Systems chemistry
RNA, Guide, CRISPR-Cas Systems genetics
Reverse Transcriptase Polymerase Chain Reaction
Severe acute respiratory syndrome-related coronavirus genetics
SARS-CoV-2 genetics
Sensitivity and Specificity
Sputum virology
Bacterial Proteins metabolism
CRISPR-Associated Proteins metabolism
Endodeoxyribonucleases metabolism
Middle East Respiratory Syndrome Coronavirus isolation & purification
Severe acute respiratory syndrome-related coronavirus isolation & purification
SARS-CoV-2 isolation & purification
Virus Diseases diagnosis
Subjects
Details
- Language :
- English
- ISSN :
- 2079-6374
- Volume :
- 11
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Biosensors
- Publication Type :
- Academic Journal
- Accession number :
- 34562891
- Full Text :
- https://doi.org/10.3390/bios11090301