한빛사 논문
Chang Ha Woo 1,5, Sungho Jang 2,3,4,5, Giyoung Shin1, Gyoo Yeol Jung 1,2,* and Jeong Wook Lee 1,2,*
1School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Republic of Korea.
2Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Republic of Korea.
3Present address: Department of Bioengineering and Nano-Bioengineering, Incheon National University, Incheon, Republic of Korea.
4Present address: Division of Bioengineering, Incheon National University, Incheon, Republic of Korea.
5These authors contributed equally: Chang Ha Woo, Sungho Jang.
*Corresponding author
Abstract
The control of viral outbreaks requires nucleic acid diagnostic tests that are sensitive, simple and fast. Here, we report a highly sensitive and specific one-pot assay for the fluorescence-based detection of RNA from pathogens. The assay, which can be performed within 30–50 min of incubation time and can reach a limit of detection of 0.1-attomolar RNA concentration, relies on a sustained isothermal reaction cascade producing an RNA aptamer that binds to a fluorogenic dye. The RNA aptamer is transcribed by the T7 RNA polymerase from the ligation product of a promoter DNA probe and a reporter DNA probe that hybridize with the target single-stranded RNA sequence via the SplintR ligase (a Chlorella virus DNA ligase). In 40 nasopharyngeal SARS-CoV-2 samples, the assay reached positive and negative predictive values of 95 and 100%, respectively. We also show that the assay can rapidly detect a range of viral and bacterial RNAs.
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TOP52020년 선정
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