文献类型：期刊文献

中文题名：基于纳米金与纳米银簇间表面等离子增强能量转移效应特异性检测microRNA

英文题名：Specific microRNA Detection Based on Surface Plasmon-Enhanced Energy Transfer Between Gold Nanoparticles and Silver Nanoclusters

作者：王红亚[1,2];尹斌成[2];叶邦策[1,2]

机构：[1]石河子大学化学和化学工程学院,石河子832000;[2]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2017

卷号：45

期号：12

起止页码：2018

中文期刊名：分析化学

外文期刊名：Chinese Journal of Analytical Chemistry

收录：CSTPCD;;Scopus;WOS:【SCI-EXPANDED(收录号:WOS:000419147100025)】；北大核心:【北大核心2014】；CSCD:【CSCD2017_2018】；

基金：This work was supported by the National Natural Science Foundation of China (Nos. 21675052, 21335003, 21575089) and the Fundamental Research Funds for the Central Universities, China.

语种：中文

中文关键词：microRNA检测;纳米金;纳米银簇;表面等离子增强能量转移效应

外文关键词：MicroRNA detection;Gold nanoparticles;Silver nanoclusters;Surface plasmon-enhanced energy transfer

摘要：microRNAs(miRNAs)的灵敏检测对临床诊断具有十分重要的意义。本研究采用偶联DNA聚合酶和核酸内切酶介导的恒温扩增反应实现靶标循环再生的策略,利用纳米金(Au NPs)与纳米银簇(Ag NCs)间表面等离子增强能量转移效应,开发了一种miRNA定量检测方法。在AuNPs表面组装两种探针(Probe a和Probe b)制备响应元件Probe b-Probe a-Au NP,其中Probe a通过3'端巯基共价偶联到Au NPs表面,此外具有靶标miRNA互补序列、核酸内切酶酶切序列和Probe b互补序列,Probe b为荧光Ag NCs合成模板。靶标miRNA存在时,启动酶级联恒温扩增反应,导致Probe b脱离Au NPs表面,抑制了Probe b为模板合成的Ag NCs与Au NPs间表面等离子增强能量转移效应,使得反应体系荧光信号增强。本方法的检出限为2.5×10^(-1)1mol/L,与miRNAs商业化检测试剂盒相比,避免了逆转录反应,而且操作简单,检测成本低,可应用于生物样本中miRNAs分析。
There is high demand for a sensitive method for miRNA detection in clinical diagnosis. In this work,we developed a method for miRNA detection based on the surface plasmon-enhanced energy transfer( SPEET) between gold nanoparticles( Au NPs) and silver nanoclusters( Ag NCs),coupled with DNA polymerase and nicking enzyme-assisted isothermal amplification for target recycling. Two DNA probes( Probe a and Probe b) were assembled onto the surface of Au NPs to form Probe b-Probe a-Au NP conjugates. Probe a consisted three domains: the complementary sequence of miRNA,the specific site of the nicking enzyme,and the self-assembly sequence for Ag NCs. The 3' end of Probe a was modified with thiol as a binding site for Au NPs. The SPEET of Ag NCs and Au NPs was inhibited when miRNA was added to produce the dumbbell shaped template by polymerase. The template could promote synthesis of Ag NCs,resulting in replacement and subsequently recycling of the target molecule for signal amplification. In comparison with the traditional method of miRNA detection with commercial RT-PCR kits,this method avoided the process of reverse transcription and was easy to perform. In addition,this method with a detection limit of 2.5×10^(-11) mol/L was cost-effective,label-free,and highly selective for detecting miRNA,and could be applied to the analysis of miRNA in biological samples.