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

Tingting Fan; Yu Mao; Feng Liu; Wei Zhang; Jin-Shun Lin; Jingxian Yin; Ying Tan; Xiaoting Huang; Yuyang Jiang

doi:10.1016/j.talanta.2019.01.038

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

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

Authors

Tingting Fan¹, Yu Mao², Feng Liu³, Wei Zhang³, Jin-Shun Lin³, Jingxian Yin¹, Ying Tan³, Xiaoting Huang¹, Yuyang Jiang⁴

Affiliations

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.

PMID: 31036212
DOI: 10.1016/j.talanta.2019.01.038

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.

MeSH terms

Biosensing Techniques*
Circulating MicroRNA / blood*
Circulating MicroRNA / isolation & purification
Endonucleases / chemistry*
Fluorescence*
Fluorescent Dyes / chemical synthesis
Fluorescent Dyes / chemistry
Humans
Nucleic Acid Amplification Techniques*
Tumor Cells, Cultured

Substances

Circulating MicroRNA
Fluorescent Dyes
Endonucleases