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10.3389-fbioe.2022.831332 |
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|a 22964185 (ISSN)
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|a Highly Sensitive Detection Method for HV69-70del in SARS-CoV-2 Alpha and Omicron Variants Based on CRISPR/Cas13a
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|b Frontiers Media S.A.
|c 2022
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|z View Fulltext in Publisher
|u https://doi.org/10.3389/fbioe.2022.831332
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|a As SARS-CoV-2 variants continue to evolve, identifying variants with adaptive diagnostic tool is critical to containing the ongoing COVID-19 pandemic. Herein, we establish a highly sensitive and portable on-site detection method for the HV69-70del which exist in SARS-CoV-2 Alpha and Omicron variants using a PCR-based CRISPR/Cas13a detection system (PCR-CRISPR). The specific crRNA (CRISPR RNA) targeting the HV69-70del is screened using the fluorescence-based CRISPR assay, and the sensitivity and specificity of this method are evaluated using diluted nucleic acids of SARS-CoV-2 variants and other pathogens. The results show that the PCR-CRISPR detection method can detect 1 copies/μL SARS-CoV-2 HV69-70del mutant RNA and identify 0.1% of mutant RNA in mixed samples, which is more sensitive than the RT-qPCR based commercial SARS-CoV-2 variants detection kits and sanger sequencing. And it has no cross reactivity with ten other pathogens nucleic acids. Additionally, by combined with our previously developed ERASE (Easy-Readout and Sensitive Enhanced) lateral flow strip suitable for CRISPR detection, we provide a novel diagnosis tool to identify SARS-CoV-2 variants in primary and resource-limited medical institutions without professional and expensive fluorescent detector. Copyright © 2022 Niu, Han, Dong, Yang, Li, Zhang, Hu, Xia, Li and Sun.
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|a Biomolecules
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|a CRISPR/cas13a
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|a CRISPR/Cas13a
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|a Detection methods
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|a Detection system
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|a Diagnosis
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|a Diagnostics tools
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|a Diseases
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|a Fluorescence
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|a Highly sensitive detections
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|a Lateral Flow
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|a lateral flow strip
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|a Lateral flow strip
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|a nucleic acid detection
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|a On-site detection
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|a Polymerase chain reaction
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|a RNA
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|a SARS
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|a SARS-CoV-2
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|a Variant
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|a variants
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|a Dong, X.
|e author
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|a Han, Y.
|e author
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|a Hu, Q.
|e author
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|a Li, F.
|e author
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|a Li, H.
|e author
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|a Niu, M.
|e author
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|a Sun, Y.
|e author
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|a Xia, X.
|e author
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|a Yang, L.
|e author
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|a Zhang, Y.
|e author
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|t Frontiers in Bioengineering and Biotechnology
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