期刊
NATURE BIOMEDICAL ENGINEERING
卷 4, 期 12, 页码 1168-1179出版社
NATURE PORTFOLIO
DOI: 10.1038/s41551-020-00617-5
关键词
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资金
- C1 Gas Refinery Program through the National Research Foundation of Korea (NRF) - Ministry of Science and ICT [NRF-2015M3D3A1A01064926, NRF-2018M3D3A1A01055754]
- NRF - Korean government (Ministry of Science and ICT) [2018R1C1B3007409, 2019R1A2C2008680]
- Human Resources Program in Energy Technology of the Korea Institute of Energy Technology Evaluation and Planning (KETEP) from the Ministry of Trade, Industry and Energy, Republic of Korea [20194030202330]
- National Research Foundation of Korea [2019R1A2C2008680, 4120200413631, 2018R1C1B3007409] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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 (aChlorellavirus 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. A one-pot enzymatic assay for the fluorescence detection of RNA accurately and rapidly detects specific viral and bacterial pathogens, as shown for SARS-CoV-2 RNA in clinical samples.
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