文献类型：期刊文献

英文题名：Colorimetric detection of sequence-specific microRNA based on duplex-specific nuclease-assisted nanoparticle amplification

作者：Wang, Qian[1];Li, Ru-Dong[1];Yin, Bin-Cheng[1];Ye, Bang-Ce[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Biomed Nanotechnol Ctr, Lab Biosyst & Microanal, Shanghai 200237, Peoples R China

年份：2015

卷号：140

期号：18

起止页码：6306

外文期刊名：ANALYST

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000360648500019)】；

基金：This work was jointly supported by the National Natural Science Foundation of China (Grants 21335003, 21205040), the Science Fund for Creative Research Groups (21421004), the Key Grant Project of Chinese Ministry of Education (Grant 313019), the Shanghai Fund (12ZR1442700), and the Fundamental Research Funds for the Central Universities, and Hitachi, Ltd.

语种：英文

摘要：Developing simple and rapid methods for sequence-specific microRNA (miRNA) analysis is imperative to the miRNA study and use in clinical diagnosis. We have developed a colorimetric method for miRNA detection based on duplex-specific nuclease (DSN)-assisted signal amplification coupled to the aggregation of gold nanoparticles (AuNPs). The proposed method involves two processes: target-mediated probe digestion by a DSN enzyme and probe-triggered AuNP aggregation as a switch for signal output. The reaction system consists of a rationally designed probe complex formed by two partly complementary DNA probes, and two sets of different oligonucleotide-modified AuNPs with sequences complementary to a DNA probe in the probe complex. In the presence of target miRNA, the probe complex is invaded, resulting in the formation of a miRNA-probe heteroduplex as the substrate of the DSN enzyme, and releasing the other probe to link to the AuNPs. The proposed method allows quantitative detection of miR-122 in the range of 20 pM to 1 nM with a detection limit of similar to 16 pM, and shows an excellent ability to discriminate single-base differences. Moreover, the detection assay can be applied to accurately quantify miR-122 in cancerous cell lysates which is in excellent agreement with the results from a commercial miRNA detection kit. This method is simple, cost-effective, highly selective, and free of dye label and separation procedures.