Issue 29, 2018, Issue in Progress

Rapid and label-free fluorescence bioassay for microRNA based on exonuclease III-assisted cycle amplification

Abstract

The quantitative analysis of microRNA is extremely important in biological research and clinical diagnosis due to the relationship between microRNA and disease. In this study, we reported a new assay for the rapid and simple detection of microRNA based on G-quadruplex and exonuclease III (ExoIII) dual signal amplification. We specifically designed two hairpins with G-quadruplex sequence. In the absence of a target, the G-quadruplex sequences are enclosed in the hairpin and fluorescence signal shut down. However, when a target is added, the dual cycle is carried out because two hairpins are digested and X and Y sequences are released under the action of ExoIII. Then, these released sequences form the G-quadruplex sequence, and N-methylmorpholine (NMM) is embedded in the G-quadruplex to produce strong fluorescence. The linear range is from 2.5 × 10−10 to 4 × 10−9 mol L−1 with a low detection limit of 6 pM. Compared to some of the previous strategies, this bioassay needs only a simple one-step reaction, and is easy for realizing the rapid detection of microRNAs. The time required for the entire analysis is only 1 hour. In addition, this bioassay has good specificity and can be applied to the actual samples.

Graphical abstract: Rapid and label-free fluorescence bioassay for microRNA based on exonuclease III-assisted cycle amplification

Supplementary files

Article information

Article type
Paper
Submitted
23 Feb 2018
Accepted
18 Mar 2018
First published
30 Apr 2018
This article is Open Access
Creative Commons BY-NC license

RSC Adv., 2018,8, 15967-15972

Rapid and label-free fluorescence bioassay for microRNA based on exonuclease III-assisted cycle amplification

M. X. Liu, S. Liang, Y. Tang, J. Tian, Y. Zhao and S. Zhao, RSC Adv., 2018, 8, 15967 DOI: 10.1039/C8RA01605D

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