Ultrasensitive Isothermal Detection of SARS-CoV-2 Based on Self-Priming Hairpin-Utilized Amplification of the G-Rich Sequence

Yan Li, Hansol Kim, Yong Ju, Yeonkyung Park, Taejoon Kang, Dongeun Yong, Hyun Gyu Park

Research output: Contribution to journalArticlepeer-review


The outbreak of the novel coronavirus disease 2019 (COVID-19) pandemic induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused millions of fatalities all over the world. Unquestionably, the effective and timely testing for infected individuals is the most imperative for the prevention of the ongoing pandemic. Herein, a new method was established for detecting SARS-CoV-2 based on the self-priming hairpin-utilized isothermal amplification of the G-rich sequence (SHIAG). In this strategy, the target RNA binding to the hairpin probe (HP) was uniquely devised to lead to the self-priming-mediated extension followed by the continuously repeated nicking and extension reactions, consequently generating abundant G-rich sequences from the intended reaction capable of producing fluorescence signals upon specifically interacting with thioflavin T (ThT). Based on the unique isothermal design concept, we successfully identified SARS-CoV-2 genomic RNA (gRNA) as low as 0.19 fM with excellent selectivity by applying only a single HP and further verified its practical diagnostic capability by reliably testing a total of 100 clinical specimens for COVID-19 with 100% clinical sensitivity and specificity. This study would provide notable insights into the design and evolution of new isothermal strategies for the sensitive and facile detection of SARS-CoV-2 under resource constraints.

Original languageEnglish
Pages (from-to)17448-17455
Number of pages8
JournalAnalytical Chemistry
Issue number50
Publication statusPublished - 2022 Dec 20

Bibliographical note

Funding Information:
This research was supported by the Mid-career Researcher Support Program of the National Research Foundation (NRF) funded by MSIT of Korea (NRF-2021R1A2B5B03001739) and KRIBB Research Initiative Program (1711134081). This research was also supported by the Korean National Police Agency (Project Name: Development of visualization technology for biological evidence in crime scenes based on nano-biotechnology/Project Number: PA-K000001-2019-101).

Publisher Copyright:
© 2022 American Chemical Society.

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry


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