Novel strategies for COVID-19 detection by utilizing nicking enzyme-based isothermal amplification

Abstract

We herein describe molecular diagnostic methods for the novel coronavirus disease (COVID-19) detection utilizing nicking enzyme-based isothermal amplification. The COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which was accompanied by high contagious and mortality, has developed into a pandemic that threatens the world. Because of its seriousness, early diagnosis of the COVID-19 is important not only for epidemiological profile but also for appropriate treatment before spreading out diseases. To perform rapid on-site detection without a sophisticated instrument, many kinds of isothermal nucleic acid amplification methods have been developed instead of the conventional PCR-based methods. In this study, nicking enzyme-based isothermal amplification newly developed by our group with high amplification efficiency and sensitivity was applied to various technology development and detection platforms for SARS-CoV-2 detection. These molecular diagnostic systems utilizing nicking enzyme-based isothermal amplification have successfully detected the genomic RNA (gRNA) extracted from SARS-CoV-2 through thermal cycler elimination, multi-analysis capability, and high sensitivity performance, and have shown great potential for application in clinical and point-of-care (POC) testing.