Fluorometric determination of microRNA-122 by using ExoIII-aided recycling amplification and polythymine induced formation of copper nanoparticles

被引：19

作者：

Tang, Yafang ^{[1
]}

Liu, Mingxiu ^{[1
]}

Zhao, Zilin ^{[1
]}

Li, Qing ^{[1
]}

Liang, Xuehua ^{[1
]}

Tian, Jianniao ^{[1
]}

Zhao, Shulin ^{[1
]}

机构：

[1] Guangxi Normal Univ, Key Lab Chem & Mol Engn Med Resources, Minist Educ China, Sch Chem & Pharmaceut Sci, Guilin 541004, Peoples R China

来源：

MICROCHIMICA ACTA | 2019年 / 186卷 / 03期

基金：

中国国家自然科学基金;

关键词：

MicroRNA; PolyT-CuNPs; Polymerization; Target recycling; LABEL-FREE DETECTION; TEMPLATED FORMATION; GOLD NANOPARTICLES; FLUORESCENT SENSOR; PLATFORM;

D O I：

10.1007/s00604-019-3237-8

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The authors describe a method for the determination of microRNA-122 by using terminal deoxynucleotidyl transferase (TdT). It is based on the use of polythymine and exonuclease III-aided cycling amplification. A 3-phosphorylated hairpin probe 1 (H-1) and a hairpin probe 2 (H-2) were designed. In the presence of the microRNA, hybridization and enzymatic cleavage will occur and produce lots of 3-hydroxylated ssDNA which can be tailed by TdT and converted into long polythymine (polyT) sequences. These can be used to synthesize copper nanoparticles (CuNPs) with fluorescence excitation/emission maxima at 350nm/630nm. This method shows good selectivity and high sensitivity with a linear response in the 1.00x10(2) fM and 1.00x10(6) fM microRNA concentration range and a 44 fM limit of detection. It was successfully applied to determination of microRNA in spiked serum samples.

引用

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