Enzyme-free and protein-assisted dual-amplified fluorescence anisotropy for sensitive miRNA detection in tumor cells

Chaofei Zeng; Jiafeng Gao; Yifei Lou; Liang Cui

doi:10.1016/j.talanta.2020.121179

Enzyme-free and protein-assisted dual-amplified fluorescence anisotropy for sensitive miRNA detection in tumor cells

Talanta. 2020 Oct 1:218:121179. doi: 10.1016/j.talanta.2020.121179. Epub 2020 May 23.

Authors

Chaofei Zeng¹, Jiafeng Gao¹, Yifei Lou¹, Liang Cui²

Affiliations

¹ Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou, 310008, China.
² Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou, 310008, China. Electronic address: cuiliang@hnu.edu.cn.

PMID: 32797926
DOI: 10.1016/j.talanta.2020.121179

Abstract

We here report a double amplification strategy to construct a fluorescence anisotropy sensor for microRNA analysis in practical biological samples. In this strategy, one target can trigger cyclic catalyzed hairpin assembly (CHA), with streptavidin incorporated as an ampliﬁer of molar mass to enhance the signal intensity. The proposed strategy has a good linearity in the range of 5 pM - 0.5 nM with a detection limit down to 2.3 pM. More importantly, by using fluorescence anisotropy as the signal output, the strategy can be used directly for detection of miRNA in practical samples without any tedious sample pretreatment, holding the practical value in real biological systems.

Keywords: Catalyzed hairpin assembly; Fluorescence anisotropy; Mass amplification; MicroRNA; Practical samples; Sensitivity.

MeSH terms

Biosensing Techniques*
Fluorescence Polarization
Limit of Detection
MicroRNAs* / genetics
Streptavidin

Substances

MicroRNAs
Streptavidin