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Talanta. 2019 Aug 1;200:480-486. doi: 10.1016/j.talanta.2019.01.038. Epub 2019 Jan 11.

Label-free fluorescence detection of circulating microRNAs based on duplex-specific nuclease-assisted target recycling coupled with rolling circle amplification.

Author information

1
Department of Chemistry, Tsinghua University, Beijing 100084, PR China; State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China.
2
School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, PR China. Electronic address: maoyu@hfut.edu.cn.
3
State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China.
4
Department of Chemistry, Tsinghua University, Beijing 100084, PR China; State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China. Electronic address: jiangyy@sz.tsinghua.edu.cn.

Abstract

A simpler, more rapid and selective biosensor has been developed for the sensitive detection of circulating miRNAs (c-miRNAs) based on duplex-specific nuclease (DSN)-assisted target recycling coupled with rolling circle amplification (RCA) using SYBR Gold Dye as a signal indicator. This biosensor exhibits a tremendously low detection limit of 0.3 fM of c-miRNAs and shows high selectivity for one-base mismatched c-miRNA. In addition, most importantly, the biosensor could accurately determine the amounts of target c-miRNA in total c-miRNAs isolated from human serums and human cancer cell lines, which confirms that the biosensor could be used to quantify c-miRNAs in complex biological samples and for the clinical early diagnosis of cancers based on c-miRNAs detection.

KEYWORDS:

Biosensor; Duplex specific nuclease; Rolling circle amplification; SYBR gold dye; Serum circulating microRNAs

PMID:
31036212
DOI:
10.1016/j.talanta.2019.01.038
[Indexed for MEDLINE]

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