Song Chunxia, Yang Xiaohai, Wang Kemin, Wang Qing, Huang Jin, Liu Jianbo, Liu Wei, Liu Pei
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry & Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China.
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry & Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China.
Anal Chim Acta. 2014 May 27;827:74-9. doi: 10.1016/j.aca.2014.04.006. Epub 2014 Apr 12.
A label-free and non-enzymatic amplification fluorescent method for detection of DNA has been developed by using hybridization chain reaction (HCR) and dsDNA-templated copper nanoparticles (CuNPs). First, the biotinylated capture DNA probes were immobilized on the streptavidin-modified beads through the interaction of biotin and streptavidin. Then, target DNA hybridized with the capture DNA probes, which formed a hybridized DNA with sticky end. The sticky end triggered the HCR process and formation of dsDNA polymers while two hairpin probes coexisted. Subsequently, the dsDNA polymers were employed as template for synthesis of CuNPs with excellent fluorescent properties, which provided a label-free, non-enzymatic signal response. Meanwhile, the fluorescence sensing depended on the target DNA triggered HCR, which render this method a high selectivity against single-base mismatch sequences. The concept and methodology developed in this work show great promise in the quantitative detection of DNA in biological and medical applications.
通过使用杂交链式反应（HCR）和双链DNA模板化铜纳米颗粒（CuNPs），开发了一种用于检测DNA的无标记非酶扩增荧光方法。首先，通过生物素与链霉亲和素的相互作用，将生物素化的捕获DNA探针固定在链霉亲和素修饰的珠子上。然后，目标DNA与捕获DNA探针杂交，形成具有粘性末端的杂交DNA。当两种发夹探针共存时，粘性末端触发HCR过程并形成双链DNA聚合物。随后，双链DNA聚合物被用作模板合成具有优异荧光特性的铜纳米颗粒，提供了无标记的非酶信号响应。同时，荧光传感依赖于目标DNA触发的HCR，这使得该方法对单碱基错配序列具有高选择性。本工作中开发的概念和方法在生物和医学应用中DNA的定量检测方面显示出巨大的潜力。
