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RESEARCH PRODUCT
CRISPR-Cas12a-Based Detection of SARS-CoV-2 Harboring the E484K Mutation
Roser Montagud-martínezSandra AlbertPilar Domingo-calapMaría-carmen MarquesRaúl RuizGuillermo RodrigoRosa Márquez-costasubject
PolymersBiomedical EngineeringBiosensing TechniquesBiologyBiochemistry Genetics and Molecular Biology (miscellaneous)Genomechemistry.chemical_compoundCOVID-19 TestingPeptide LibraryTechnical NoteCRISPRCRISPR diagnosticsHumansGeneticsvirus evolutionSARS-CoV-2Epidemiological surveillanceepidemiological surveillanceCOVID-19General MedicineDNAAmpliconSurface Plasmon ResonanceVirus evolutionProtospacer adjacent motifHEK293 CellschemistryGenetic TechniquesSpainViral evolutionImmunoglobulin GMutation (genetic algorithm)DNA ViralMutationRespiratory virusCRISPR-Cas SystemsDNAdescription
The novel respiratory virus SARS-CoV-2 is rapidly evolving across the world with the potential of increasing its transmission and the induced disease. Here, we applied the CRISPR-Cas12a system to detect, without the need of sequencing, SARS-CoV-2 genomes harboring the E484K mutation, first identified in the Beta variant and catalogued as an escape mutation. The E484K mutation creates a canonical protospacer adjacent motif for Cas12a recognition in the resulting DNA amplicon, which was exploited to obtain a differential readout. We analyzed a series of fecal samples from hospitalized patients in Valencia (Spain), finding one infection with SARS-CoV-2 harboring the E484K mutation, which was then confirmed by sequencing. Overall, these results suggest that CRISPR diagnostics can be a useful tool in epidemiology to monitor the spread of escape mutations
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2021-11-16 | ACS Synthetic Biology |