Simultaneous Dual‐Gene Diagnosis of SARS‐CoV‐2 Based on CRISPR/Cas9‐Mediated Lateral Flow Assay

Few methods for the detection of SARS-CoV-2 currently have capability to simultaneously detect two genes in a single test, which is key measure improve accuracy, as adopted by gold standard RT-qPCR method. Developed here CRISPR/Cas9-mediated triple-line lateral flow assay (TL-LFA) combined with multiplex reverse transcription-recombinase polymerase amplification (RT-RPA) rapid and simultaneous dual-gene strip test. This characterized envelope (E) open reading frame 1ab (Orf1ab) from cell-cultured viral RNA standards, showing sensitivity 100 copies per reaction (25 μL). Furthermore, analysis 64 nasopharyngeal swab samples showed % negative predictive agreement 97.14 positive agreement. platform will provide more accurate convenient pathway diagnosis COVID-19 or other infectious diseases low-resource regions.