Department of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea.
School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea.
Int J Mol Sci. 2021 Feb 10;22(4):1772. doi: 10.3390/ijms22041772.
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.
需要一种快速、灵敏且简单的即时（POC）核酸诊断测试来防止传染病的传播。基于纸的引发反应是一种新兴的比色 POC 核酸诊断测试，已被广泛用于病原体检测和微生物组分析。在这里，我们在引发反应之前引入了一种称为逆转录环介导扩增（RT-LAMP）的扩增方法，并对其进行了修改，以实现对 RNA 病毒更灵敏和更快的比色检测。我们表明，将改良的 RT-LAMP 与引发反应结合使用，可以在 70 分钟内检测到低至 120 个拷贝的冠状病毒 RNA。针对其他冠状病毒的交叉反应测试表明，这种带有改良 RT-LAMP 的引发反应对靶 RNA 具有高度特异性。总的来说，带有改良 RT-LAMP 的基于纸的引发开关传感器可实现快速、灵敏、特异性和比色冠状病毒检测。
