期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 60, 期 10, 页码 5307-5315出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202014506
关键词
genes; polymerases; RNA; SARS-CoV-2; viruses
资金
- National Natural Science Foundation of China [91959128, 21874049, 21904042, 81772246]
- Special Project of Science and Technology Development of Guangdong Province [2017B020207011]
- Special Support Program of Guangdong Province [2016TQ03R749]
- State Key Laboratory of Chemo/Biosensing and Chemometrics of Hunan University [2019004]
- State Key Laboratory of Electroanalytical Chemistry [SKLEAC202001]
- Key Research and Development Plan of Hubei Province [2020BCA090]
The newly developed method can simultaneously detect two SARS-CoV-2 genes in a single test, showing high sensitivity and accuracy, providing a more accurate and convenient pathway for COVID-19 diagnosis in low-resource regions.
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 mu L). 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.
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