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Simultaneous Dual‐Gene Diagnosis of SARS‐CoV‐2 Based on CRISPR/Cas9‐Mediated Lateral Flow Assay
Author(s) -
Xiong Erhu,
Jiang Ling,
Tian Tian,
Hu Menglu,
Yue Huahua,
Huang Mengqi,
Lin Wei,
Jiang Yongzhong,
Zhu Debin,
Zhou Xiaoming
Publication year - 2021
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202014506
Subject(s) - crispr , recombinase polymerase amplification , multiplex , cas9 , gene , gold standard (test) , virology , biology , microbiology and biotechnology , computational biology , polymerase chain reaction , genetics , medicine
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.