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Sensors 2017, 17(4), 760; doi:10.3390/s17040760

An Exonuclease I-Based Quencher-Free Fluorescent Method Using DNA Hairpin Probes for Rapid Detection of MicroRNA

State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China
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Received: 3 February 2017 / Revised: 26 March 2017 / Accepted: 31 March 2017 / Published: 3 April 2017
(This article belongs to the Special Issue Genosensing)
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Abstract

MicroRNAs (miRNAs) act as biomarkers for the diagnosis of a variety of cancers. Since the currently used methods for miRNA detection have limitations, simple, sensitive, and cost-effective methods for the detection of miRNA are required. This work demonstrates a facile, quencher-free, fluorescence-based analytical method for cost-effective and sensitive detection of miRNA using a super 2-aminopurine (2-AP)-labeled hairpin probe (HP) and exonuclease I activity. Specifically, the fluorescence of 2-AP is strongly quenched when it is incorporated within DNA. In the presence of a target miRNA, HP attains an open conformation by hybridizing with the target miRNA to form a double-stranded structure with a protruding 3′-terminus. Next, the digestion of the protruding 3′-terminus is triggered by exonuclease I, during which 2-AP is released free in solution from the DNA, thereby increasing fluorescence. This method is highly sensitive, with a detection limit of 0.5 nM—10 times lower than a previously reported quencher-free fluorescence method. Furthermore, this method has potential applications in clinical diagnosis and biomedical research. View Full-Text
Keywords: microRNA; quencher-free; exonuclease I; 2-aminopurine; hairpin probe microRNA; quencher-free; exonuclease I; 2-aminopurine; hairpin probe
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MDPI and ACS Style

Ma, C.; Liu, H.; Wu, K.; Chen, M.; Zheng, L.; Wang, J. An Exonuclease I-Based Quencher-Free Fluorescent Method Using DNA Hairpin Probes for Rapid Detection of MicroRNA. Sensors 2017, 17, 760.

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