Park S(1), Lee JW(1)(2). Author information:
(1)Department of Chemical Engineering, Pohang University of Science and
Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea.
(2)School of Interdisciplinary Bioscience and Bioengineering, Pohang University
of Science, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea.
A rapid, sensitive and simple point-of-care (POC) nucleic acid diagnostic test is needed to prevent spread of infectious diseases. Paper-based toehold reaction, a recently emerged colorimetric POC nucleic acid diagnostic test, has been widely used for pathogen detection and microbiome profiling. Here, we introduce an amplification method called reverse transcription loop-mediated amplification (RT-LAMP) prior to the toehold reaction and modify it to enable more sensitive and faster colorimetric detection of RNA viruses. We show that incorporating the modified RT-LAMP to the toehold reaction detects as few as 120 copies of coronavirus RNA in 70 min. Cross-reactivity test against other coronaviruses indicates this toehold reaction with the modified RT-LAMP is highly specific to the target RNA. Overall, the paper-based toehold switch sensors with the modified RT-LAMP allow fast, sensitive, specific and colorimetric coronavirus detection.
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