Direct labeling microRNA with an electrocatalytic moiety and its application in ultrasensitive microRNA assays

被引：76

作者：

Gao, Zhiqiang

Yu, Yuan Hong

机构：

[1] ASTAR, Inst Microelect, Singapore 117685, Singapore

[2] ASTAR, Ctr Mol Med, Singapore 138673, Singapore

来源：

BIOSENSORS & BIOELECTRONICS | 2007年 / 22卷 / 06期

关键词：

microRNA; electrocatalysis; amperometry; 1,10-phenanthroline-5,6-dione; ruthenium;

D O I：

10.1016/j.bios.2006.04.020

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

An ultrasensitive procedure for the detection of microRNA (miRNA) in total RNA is described in this work. The miRNA is directly labeled with a redox active and electrocatalytic moiety, Ru(PD)(2)Cl(2) (PD = 1, 10-phenanthroline-5,6-dione), through coordinative bonds with purine bases in the miRNA molecule. The excellent electrocatalytic activity of the Ru(PD)2Cl(2) towards the oxidation of hydrazine makes it possible to conduct ultrasensitive miRNA detection. Under optimized experimental conditions, the assay allows the detection of miRNAs in the range of 0.50-400 pM with a detection limit of 0.20 pM in 2.5 mu l (0.50 amole). MicroRNA quantitation is therefore performed in as little as 10 ng of total RNA, providing a much-needed platform for miRNA expression analysis. (c) 2006 Published by Elsevier B.V.

引用

收藏

页码：933 / 940

页数：8

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