Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun, 130012 China.
Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun, 130012 China.
Biosens Bioelectron. 2017 Jan 15;87:66-72. doi: 10.1016/j.bios.2016.07.061. Epub 2016 Jul 20.
In this work, a novel fluorescent DNA sensor for ultrasensitive detection of Helicobacter pylori (H. pylori) DNA was developed. This strategy took advantage of DNA hybridization between single-stranded DNA (ssDNA, which had been designed as an aptamer specific for H. pylori DNA) and the complementary target H. pylori DNA, and the feature that ssDNA bound to graphene oxide (GO) with significantly higher affinity than double-stranded DNA (dsDNA). ssDNA were firstly covalent conjugated with CuInS quantum dots (QDs) by reaction between the carboxy group of QDs and amino group modified ssDNA, forming ssDNA-QDs genosensor. In the absence of the complementary target H. pylori DNA, GO could adsorb ssDNA-QDs DNA sensor and efficiently quench the fluorescence of ssDNA-QDs. While the complementary target H. pylori DNA was introduced, the ssDNA-QDs preferentially bound with the H. pylori DNA. The formation of dsDNA would alter the conformation of ssDNA and disturb the interaction between ssDNA and GO. Thus, the dsDNA-QDs/GO system exhibited a stronger fluorescence emission than that of the ssDNA-QDs/GO system. Under the optimized conditions, a linear correlation was established between the fluorescence intensity ratio I/I and the concentration of H. pylori DNA in the range of 1.25-875pmolL with a detection limit of 0.46pmolL. The proposed method was applied to the determination of H. pylori DNA sequence in milk samples with satisfactory results.
在这项工作中,开发了一种用于超灵敏检测幽门螺杆菌 (H. pylori) DNA 的新型荧光 DNA 传感器。该策略利用单链 DNA(ssDNA,其被设计为特异性识别 H. pylori DNA 的适体)与互补靶标 H. pylori DNA 之间的 DNA 杂交,以及 ssDNA 与氧化石墨烯 (GO) 结合的特性,其亲和力明显高于双链 DNA (dsDNA)。ssDNA 首先通过 QDs 上的羧基和修饰后的 ssDNA 上的氨基之间的反应与 CuInS 量子点 (QDs) 共价共轭,形成 ssDNA-QDs 基因传感器。在不存在互补靶标 H. pylori DNA 的情况下,GO 可以吸附 ssDNA-QDs DNA 传感器并有效猝灭 ssDNA-QDs 的荧光。当引入互补的靶标 H. pylori DNA 时,ssDNA-QDs 优先与 H. pylori DNA 结合。dsDNA 的形成会改变 ssDNA 的构象并干扰 ssDNA 和 GO 之间的相互作用。因此,dsDNA-QDs/GO 系统的荧光发射强度比 ssDNA-QDs/GO 系统更强。在优化条件下,荧光强度比 I/I 与 H. pylori DNA 浓度在 1.25-875pmolL 范围内呈线性相关,检测限为 0.46pmolL。该方法被应用于牛奶样品中 H. pylori DNA 序列的测定,结果令人满意。