Residential College | false |
Status | 已發表Published |
SARS-CoV-2 RNA detection with duplex-specific nuclease signal amplification | |
Liu,Meiqing1; Li,Haoran1,2; Jia,Yanwei1,2,3![]() ![]() ![]() ![]() | |
2021-02-01 | |
Source Publication | Micromachines
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ISSN | 2072-666X |
Volume | 12Issue:2 |
Abstract | The emergence of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a zoonotic pathogen, has led to the outbreak of coronavirus disease 2019 (COVID-19) pandemic and brought serious threats to public health worldwide. The gold standard method for SARSCoV- 2 detection requires both reverse transcription (RT) of the virus RNA to cDNA and then polymerase chain reaction (PCR) for the cDNA amplification, which involves multiple enzymes, multiple reactions and a complicated assay optimization process. Here, we developed a duplexspecific nuclease (DSN)-based signal amplification method for SARS-CoV-2 detection directly from the virus RNA utilizing two specific DNA probes. These specific DNA probes can hybridize to the target RNA at different locations in the nucleocapsid protein gene (N gene) of SARS-CoV-2 to form a DNA/RNA heteroduplex. DSN cleaves the DNA probe to release fluorescence, while leaving the RNA strand intact to be bound to another available probe molecule for further cleavage and fluorescent signal amplification. The optimized DSN amount, incubation temperature and incubation time were investigated in this work. Proof-of-principle SARS-CoV-2 detection was demonstrated with a detection sensitivity of 500 pM virus RNA. This simple, rapid, and direct RNA detection method is expected to provide a complementary method for the detection of viruses mutated at the PCR primer-binding regions for a more precise detection. |
Keyword | Dna Probe Duplex-specific Nuclease Rna Detection Sars-cov-2 Signal Amplification |
DOI | 10.3390/mi12020197 |
URL | View the original |
Indexed By | SCIE |
Language | 英語English |
WOS Research Area | Chemistry ; Science & Technology - Other Topics ; Instruments & Instrumentation ; Physics |
WOS Subject | Chemistry, Analytical ; Nanoscience & Nanotechnology ; Instruments & Instrumentation ; Physics, Applied |
WOS ID | WOS:000622811600001 |
Scopus ID | 2-s2.0-85101285690 |
Fulltext Access | |
Citation statistics | |
Document Type | Journal article |
Collection | DEPARTMENT OF BIOMEDICAL SCIENCES Faculty of Science and Technology THE STATE KEY LABORATORY OF ANALOG AND MIXED-SIGNAL VLSI (UNIVERSITY OF MACAU) INSTITUTE OF MICROELECTRONICS DEPARTMENT OF ELECTRICAL AND COMPUTER ENGINEERING |
Corresponding Author | Jia,Yanwei |
Affiliation | 1.State-Key Laboratory of Analog and Mixed-Signal VLSI,Institute of Microelectronics,University of Macau,Macau,999078,China 2.Faculty of Science and Technology-ECE,University of Macau,Macau,999078,China 3.Faculty of Health Sciences,University of Macau,Macau,999078,China 4.On Leave from Instituto Superior Técnico,Universidade de Lisboa,Lisboa,1049-001,Portugal |
First Author Affilication | University of Macau |
Corresponding Author Affilication | University of Macau; Faculty of Science and Technology; Faculty of Health Sciences |
Recommended Citation GB/T 7714 | Liu,Meiqing,Li,Haoran,Jia,Yanwei,et al. SARS-CoV-2 RNA detection with duplex-specific nuclease signal amplification[J]. Micromachines, 2021, 12(2). |
APA | Liu,Meiqing., Li,Haoran., Jia,Yanwei., Mak,Pui In., & Martins,Rui P. (2021). SARS-CoV-2 RNA detection with duplex-specific nuclease signal amplification. Micromachines, 12(2). |
MLA | Liu,Meiqing,et al."SARS-CoV-2 RNA detection with duplex-specific nuclease signal amplification".Micromachines 12.2(2021). |
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