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A rapid room-temperature DNA amplification and detection strategy based on nicking endonuclease and catalyzed hairpin assembly

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Abstract

A novel and rapid room-temperature DNA amplification strategy to simultaneously amplify and detect target HIV DNA was developed by coupling catalyzed hairpin assembly (CHA) with a “non-sequence dependent” nicking endonuclease. In this system, a Nt.BsmAI nicking endonuclease recognition sequence and HIV target DNA sequence were integrated into hairpin probes H1 and H3, respectively. The HIV target DNA was repeatedly used to trigger a CHA reaction, forming numerous H1–H2 duplexes. Then the newly formed sticky ends of H1–H2 caused the hairpin structure of H3 to open. As a result, the double-stranded nicking enzyme recognition site was reunited, which was recognized and cleaved by the nicking enzyme. Finally, the target DNA replica with a sequence matching that of the HIV target DNA was obtained. The above reactions, namely, the CHA reaction, hairpin opening and enzyme cleavage, can be continuously repeated, leading to rapid room-temperature DNA amplification. In addition, this proposed rapid DNA amplification strategy enables picomolar detection of HIV DNA target within 10 min, providing a potential clinical application at point-of-care.

Graphical abstract: A rapid room-temperature DNA amplification and detection strategy based on nicking endonuclease and catalyzed hairpin assembly

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Publication details

The article was received on 11 Mar 2019, accepted on 06 Apr 2019 and first published on 09 Apr 2019


Article type: Paper
DOI: 10.1039/C9AY00507B
Citation: Anal. Methods, 2019, Advance Article

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    A rapid room-temperature DNA amplification and detection strategy based on nicking endonuclease and catalyzed hairpin assembly

    G. Dong, J. Dai, L. Jin, H. Shi, F. Wang, C. Zhou, B. Zheng, Y. Guo and D. Xiao, Anal. Methods, 2019, Advance Article , DOI: 10.1039/C9AY00507B

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