Chip-based visual detection of micro RNA using DNA-functionalized gold nanoparticles  

作　　者：Ping Wang Jianlong Zhao Bin Hu Zule Cheng Yanan Bai Qinghui Jin Huiying Liu Hongju Mao Sanqiang Li Jianlong Zhao 

机构地区：[1]The Molecular Medicine Key Laboratory of Liver Injury and Repair, Medical College, Henan University of Science and Technology [2]State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences [3]Clinical Research Center of Biological Sensing Technology, Shanghai Xuhui Central Hospital [4]University of Chinese Academy of Sciences [5]Department of Oral and Maxillofacial, School of Stomatology, Dalian Medical University

出　　处：《Science China(Life Sciences)》2016年第5期510-515,共6页中国科学（生命科学英文版）

基　　金：supported by the National Basic Research Program of China (2012CB933303);the National Natural Science Foundation of China (61571429, 61571077, 61401442);the Innovation Team of Henan University of Science and Technology (2015XTD003);the Science and Technology Commission of Shanghai Municipality (12441902600, 1402H233900);the Shanghai Clinical Center/Shanghai Xuhui Central Hospital, Chinese Academic of Sciences (BRC2012002)

摘　　要：In the present study, we developed a highly sensitive and convenient biosensor consisting of gold nanoparticle （AuNP） probes and a gene chip to detect microRNAs （miRNAs）. Specific oligonucleotides were attached to the glass surface as capture probes for the target miRNAs, which were then detected via hybridization to the AuNP probes. The signal was amplified via the re- duction of HAuCI4 by H202. The use of a single AuNP probe detected 10 pmol L-1 of target miRNA. The recovery rate for miR-126 from fetal bovine serum was 81.5%-109.1%. The biosensor detection of miR-126 in total RNA extracted from lung cancer tissues was consistent with the quantitative PCR （qPCR） results. The use of two AuNP probes further improved the de- tection sensitivity such that even 1 fmol L-t of target miR-125a-5p was detectable. This assay takes less than 1 h to complete and the results can be observed by the naked eye, The platform simultaneously detected lung cancer related miR-126 and miR-125a-5p. Therefore, this low cost, rapid, and convenient technology could be used for ultrasensitive and robust visual miRNA detection.In the present study, we developed a highly sensitive and convenient biosensor consisting of gold nanoparticle(Au NP) probes and a gene chip to detect micro RNAs(mi RNAs). Specific oligonucleotides were attached to the glass surface as capture probes for the target mi RNAs, which were then detected via hybridization to the Au NP probes. The signal was amplified via the reduction of HAu Cl4 by H2O2. The use of a single Au NP probe detected 10 pmol L?1 of target mi RNA. The recovery rate for mi R-126 from fetal bovine serum was 81.5%–109.1%. The biosensor detection of mi R-126 in total RNA extracted from lung cancer tissues was consistent with the quantitative PCR(q PCR) results. The use of two Au NP probes further improved the detection sensitivity such that even 1 fmol L?1 of target mi R-125a-5p was detectable. This assay takes less than 1 h to complete and the results can be observed by the naked eye. The platform simultaneously detected lung cancer related mi R-126 and mi R-125a-5p. Therefore, this low cost, rapid, and convenient technology could be used for ultrasensitive and robust visual mi RNA detection.

关 键 词：MICRORNA gold nanoparticles CHIP BIOSENSOR visual detection 

分 类 号：TP212.3[自动化与计算机技术—检测技术与自动化装置] TB383.1[自动化与计算机技术—控制科学与工程] R440[一般工业技术—材料科学与工程]