Sensitive and enzyme-free detection for single nucleotide polymorphism using microbead-assisted toehold-mediated strand displacement reaction

作     者：Jia-Bao Long Ying-Xin Liu Qing-Feng Cao Qiu-Ping Guo Shu-Ya Yan Xiang-Xian Meng 

作者机构：College of Biology State Key Laboratory of Chemo/Biosensing and Chemometrics Hunan University Department of Bioengineering and Environment Science Changsha University 

出 版 物：《Chinese Chemical Letters》 (中国化学快报（英文版）)

年 卷 期：2015年第26卷第8期

页      面：1031-1035页

核心收录：

学科分类：081704[工学-应用化学] 07[理学] 080202[工学-机械电子工程] 08[工学] 0817[工学-化学工程与技术] 070302[理学-分析化学] 0802[工学-机械工程] 0703[理学-化学] 

基　　金：supported by National Natural Science Foundation of China(No.21275043) National Basic Research Program of China under Grants(No.2009CB421601) 

主　　题：strand capture biosensor convenient detecting capable inexpensive reporter bright matched 

摘      要：This report described a free-enzyme, convenient and inexpensive genotyping biosensor capable of detecting single nucleotide polymorphism at normal temperature based on the combination of toeholdmediated strand displacement reaction（toehold-SDR） and microbead-capture technique. The biosensor consists of a pre-hybridized strand formed by a reporter probe and a capture probe. In the presence of a mutant sequence, there is no toehold-mediated strand displacement and the reporter probe cannot be released from the pre-hybridized strand. Microbeads capture the fluorescent pre-hybridized strand through biotin–streptavidin interaction, so microbeads give out significant fluorescence signal, while there is no fluorescence in the solution. However, in the presence of a matched target, the strand displacement is effectively initiated and the reporter probe is released from pre-hybridized strand. After adding microbeads, the solution produces bright fluorescence, while microbeads have no obvious *** are identified conveniently according to the fluorescence intensity of the solution. The method provides a simple and inexpensive strategy to detect point mutation. Moreover, this biosensor shows the linear relationship in the range of 1–40 nmol/L and reaches a detection limit of 0.3 nmol/L.？ 2015 Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical *** by Elsevier B.V. All rights reserved.