Ultrasensitive and amplification-free detection of SARS-CoV-2 RNA using an electrochemical biosensor powered by CRISPR/Cas13a
- Authors
- Kashefi-Kheyrabadi, Leila; Nguyen, Huynh Vu; Go, Anna; Lee, Min-Ho
- Issue Date
- Apr-2023
- Publisher
- Elsevier B.V.
- Keywords
- CRISPR; Electrochemical biosensor; Orf1ab gene; S gene; SARS-CoV-2 RNA
- Citation
- Bioelectrochemistry, v.150
- Journal Title
- Bioelectrochemistry
- Volume
- 150
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/60614
- DOI
- 10.1016/j.bioelechem.2023.108364
- ISSN
- 1567-5394
1878-562X
- Abstract
- This study proposed a CRISPR/Cas13a-powered electrochemical multiplexed biosensor for detecting SARS-CoV-2 RNA strands. Current SARS-CoV-2 diagnostic methods, such as reverse transcription PCR (RT-PCR), are primarily based on nucleic acid amplification (NAA) and reverse transcription (RT) processes, which have been linked to significant issues such as cross-contamination and long turnaround times. Using a CRISPR/Cas13a system integrated onto an electrochemical biosensor, we present a multiplexed and NAA-free strategy for detecting SARS-CoV-2 RNA fragments. SARS-CoV-2 S and Orf1ab genes were detected in both synthetic and clinical samples. The CRISPR/Cas13a-powered biosensor achieved low detection limits of 2.5 and 4.5 ag/µL for the S and Orf1ab genes, respectively, successfully meeting the sensitivity requirement. Furthermore, the biosensor's specificity, simplicity, and universality may position it as a potential rival to RT-PCR. © 2023 Elsevier B.V.
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