Simultaneous Dual-Gene Diagnosis of SARS-CoV-2 Based on CRISPR/Cas9-Mediated Lateral Flow Assay.
Erhu XiongLing JiangTian TianMenglu HuHuahua YueMengqi HuangWei LinYongzhong JiangDebin ZhuXiaoming ZhouPublished in: Angewandte Chemie (International ed. in English) (2021)
Few methods for the detection of SARS-CoV-2 currently have the capability to simultaneously detect two genes in a single test, which is a key measure to improve detection accuracy, as adopted by the gold standard RT-qPCR method. Developed here is a CRISPR/Cas9-mediated triple-line lateral flow assay (TL-LFA) combined with multiplex reverse transcription-recombinase polymerase amplification (RT-RPA) for rapid and simultaneous dual-gene detection of SARS-CoV-2 in a single strip test. This assay is characterized by the detection of envelope (E) and open reading frame 1ab (Orf1ab) genes from cell-cultured SARS-CoV-2 and SARS-CoV-2 viral RNA standards, showing a sensitivity of 100 RNA copies per reaction (25 μL). Furthermore, dual-gene analysis of 64 nasopharyngeal swab samples showed 100 % negative predictive agreement and 97.14 % positive predictive agreement. This platform will provide a more accurate and convenient pathway for diagnosis of COVID-19 or other infectious diseases in low-resource regions.
Keyphrases
- sars cov
- crispr cas
- loop mediated isothermal amplification
- respiratory syndrome coronavirus
- high throughput
- real time pcr
- genome wide
- genome wide identification
- label free
- genome editing
- infectious diseases
- copy number
- single cell
- genome wide analysis
- transcription factor
- stem cells
- minimally invasive
- coronavirus disease
- endothelial cells
- working memory
- bioinformatics analysis