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Sensitive Detection of MicroRNA in Complex Biological Samples by Using Two Stages DSN-Assisted Target Recycling Signal Amplification Method.

Biosens Bioelectron.. 2017-01;  87:358-364
K Zhang, K Wang, X Zhu, F Xu, M Xie. Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
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Abstract

MicroRNA (miRNA) has become an important biomarker candidate for cancer diagnosis, prognosis, and therapy. In this study, we have developed a novel fluorescence method for sensitive and specific miRNA detection via duplex specific nuclease (DSN) signal amplification and demonstrated its practical application in biological samples. Malachite green (MG) was employed as a "label-free" signal transducer since fluorescence of MG could be enhanced by 100-fold when MG were binding to a G-quadruplex structure formed within the d(G2T)13G sequence. The proposed signal amplification strategy is an integrated "biological circuit" designed to initiate a cascade of enzymatic reactions in order to detect, ... More

Keywords

MiRNA; DSN; Amplification method; Malachite green; Cell lysates