文献类型：期刊文献

英文题名：QDs-DNA nanosensor for the detection of hepatitis B virus DNA and the single-base mutants

作者：Wang, Xiang[1,2];Lou, Xinhui[1];Wang, Yi[1];Guo, Qingchuan[1];Fang, Zheng[2];Zhong, Xinhua[2];Mao, Hongju[1];Jin, Qinghui[1];Wu, Lei[1];Zhao, Hui[1];Zhao, Jianlong[1]

机构：[1]Chinese Acad Sci, Shanghai Inst Microsyst & Informat Technol, Shanghai 200050, Peoples R China;[2]E China Univ Sci & Technol, Dept Chem, Shanghai 200237, Peoples R China

年份：2010

卷号：25

期号：8

起止页码：1934

外文期刊名：BIOSENSORS & BIOELECTRONICS

收录：;EI(收录号：20101012747562);WOS:【SCI-EXPANDED(收录号:WOS:000276424200012)】；

基金：This work was supported by grants from National Science and Technology Major Project (2009ZX10004-105, 2009ZX10004-301), Science and Technology Commission of Shanghai Municipality (No. 0952nm05700) and the 973 Program of the Ministry of Science and Technology of China (Nos. 2007CB714502 and 2007CB936000).

语种：英文

外文关键词：Quantum dots (QDs); Fluorescence resonance energy transfer (FRET); Hepatitis B virus (HBV); DNA; Mutation

摘要：We report here a quantum dots-DNA (QDs-DNA) nanosensor based on fluorescence resonance energy transfer (FRET) for the detection of the target DNA and single mismatch in hepatitis B virus (HBV) gene. The proposed one-pot DNA detection method is simple, rapid and efficient due to the elimination of the washing and separation steps. In this study, the water-soluble CdSe/ZnS QDs were prepared by replacing the trioctylphosphine oxide (TOPO) on the surface of QDs with 3-mercaptopropionic acid (MPA). Subsequently, oligonucleotides were attached to the QDs surface to form functional QDs-DNA conjugates. Along with the addition of DNA targets and Cy5-modified signal DNAs into the QDs-DNA conjugates, sandwiched hybrids were formed. The resulting assembly brings the Cy5 fluorophore, the acceptor, and the QDs, the donor, into proximity, leading to fluorescence emission from the acceptor by means of FRET on illumination of the donor. In order to efficiently detect single-base mutants in HBV gene, oligonucleotide ligation assay was employed. If there existed a single-base mismatch, which could be recognized by the ligase, the detection probe was not ligated and no Cy5 emission was produced due to the lack of FRET. The feasibility of the proposed method was also demonstrated in the detection of synthetic 30-mer oliginucleotide targets derived from the HBV with a sensitivity of 4.0 nM by using a multilabel counter. The method enables a simple and efficient detection that could be potentially used for high throughput and multiplex detections of target DNA and the mutants. (c) 2010 Elsevier B.V. All rights reserved.