Three-way junction-induced isothermal amplification for nucleic acid detection

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We, herein, describe a three-way junction (3WJ)-induced isothermal amplification (ThIsAmp) reaction for target nucleic acid detection. In this strategy, target nucleic acid induces the formation of 3WJ structure by associating a specially designed ThlsAmp template and ThIsAmp primer. Upon the formation of 3WJ structure, ThIsAmp primer is subjected to continuously repeated extension and nicking reaction by the combined activities of DNA polymerase and nicking endonuclease, consequently producing a large number of trigger strands. The trigger strands then initiate two separate but interconnected pathways by binding to either 3' overhang of ThlsAmp template within the 3WJ structure or free ThIsAmp template. As a consequence, a large number of final double-stranded DNA products are produced under an isothermal condition, which can be monitored in real-time by detecting the fluorescence intensity resulting from SYBR Green I staining. Based on this principle, we successfully detected target DNA down to 78.1 aM with excellent specificity. The sophisticated design principle employed in this work would provide great insight for the development of self-operative isothermal amplifying system enabling target nucleic acid detection.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2020-01
Language
English
Article Type
Article
Citation

BIOSENSORS & BIOELECTRONICS, v.147

ISSN
0956-5663
DOI
10.1016/j.bios.2019.111762
URI
http://hdl.handle.net/10203/269808
Appears in Collection
CBE-Journal Papers(저널논문)
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