高等学校化学学报Issue(8)：1646-1651,6.DOI:10.7503/cjcu20140321
基于免标记发夹型探针和核酸外切酶Ⅲ的荧光信号放大DNA检测
Label-free Fluorescence Signal Amplifying Detection of DNA Based on Hairpin Probe and Exo Ⅲ
摘要
Abstract
A novel label-free fluorescence signal amplifying method for DNA detection was developed with high specificity and sensitivity based on a long arm hairpin nucleic acid probe and exonuclease Ⅲ( Exo Ⅲ) . Without the target DNA, the SYBR Green Ⅰ dye could be embedded into the stem of hairpin nucleic acid probe to generate strong fluorescence. While in the presence of target DNA, Exo Ⅲ could catalyze the step-wise removal of mononucleotides from 3’-OH termini of double-stranded DNA with the degradation of the hair-pin probe. After that, the target DNA was released, triggering the next cycle of exonuclease digestion reac-tion. Then the SYBR Green Ⅰ was continuously released, resulting in fluorescence intensity decreased. It rea-lized the label-free signal amplifying detection of DNA. The detection limit of this method was as low as 320 fmol/L. It can be expected to provide a novel, simple, label-free and rapid tool for DNA detection.关键词
发夹型探针/核酸外切酶Ⅲ/免标记/信号放大/DNA检测Key words
Hairpin probe/Exo Ⅲ/Label-free/Signal amplification/DNA detection分类
化学化工引用本文复制引用
郭秋平,赵下雨,谢琴,王柯敏,万俊,袁宝银,谭誉宇..基于免标记发夹型探针和核酸外切酶Ⅲ的荧光信号放大DNA检测[J].高等学校化学学报,2014,(8):1646-1651,6.基金项目
国家自然科学基金重大项目(批准号:21190044)、国家自然科学基金(批准号:21175035)、国家“九七三”计划项目(批准号:2011CB911002)、科技部国际合作专项(批准号:2010DFB30300)和湖南省科学技术项目(批准号:2013FJ4042)资助.Supported by the Major Program of the National Natural Science Foundation of China(No.21190044), the National Natural Science Founda-tion of China(No.21175035), the National Basic Research Program of China(No.2011CB911002), the International Science & Technology Cooperation Program of China(No.2010DFB30300) and Hunan Provincial Science and Technology Project of China(No.2013FJ4042) （批准号:21190044）
