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基于树枝状大分子的DNA生物传感器,用于使用金纳米颗粒修饰的报告探针DNA对DNA杂交进行高灵敏度电化学检测。

Dendrimers-based DNA biosensors for highly sensitive electrochemical detection of DNA hybridization using reporter probe DNA modified with Au nanoparticles.

作者信息

Li Guangjiu, Li Xiaolin, Wan Jun, Zhang Shusheng

机构信息

Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China.

出版信息

Biosens Bioelectron. 2009 Jul 15;24(11):3281-7. doi: 10.1016/j.bios.2009.04.022. Epub 2009 Apr 24.

Abstract

A novel and sensitive electrochemical approach for sequence-specific DNA detection based on polyamidoamine (PAMAM) and signal amplification with nanoparticles (NPs) is reported. A gold electrode was first modified with 3-mercaptopropionic acid, and then reacted with an amino-terminated polyamidoamine (PAMAM, G 4.0-NH(2)) to obtain a thin film. Single-stranded 3'-biotin end-labeled oligonucleotide was immobilized onto the film to obtain a stable recognition layer through biotin-avidin combination to detect complementary target, using signal amplification with Au NPs and Ru(NH(3))(6) as redox electroactive indicators. The properties of the avidin/PAMAM/3-mercaptopropionic acid (RSH)/Au, the characteristics of the immobilization and hybridization of DNA were studied by cyclic voltammetry (CV), difference pulse voltammetry (DPV) and electrochemical impedance spectroscopy. The dynamic detection range of the sequence-specific DNA was from 1.4 x 10(-11) to 2.7 x 10(-14) mol L(-1) and the detection limit was 1.4 x 10(-14) mol L(-1). The DNA sensor not only showed low detection limit, but also exhibited excellent selectivity against two-base mismatched DNA.

摘要

报道了一种基于聚酰胺-胺(PAMAM)和纳米颗粒(NPs)信号放大的新型灵敏电化学方法用于序列特异性DNA检测。首先用3-巯基丙酸修饰金电极,然后与氨基端聚酰胺-胺(PAMAM,G 4.0-NH(2))反应得到薄膜。将3'-生物素末端标记的单链寡核苷酸固定在该薄膜上,通过生物素-抗生物素蛋白结合获得稳定的识别层,以检测互补靶标,使用金纳米颗粒和Ru(NH(3))(6)作为氧化还原电活性指示剂进行信号放大。通过循环伏安法(CV)、差分脉冲伏安法(DPV)和电化学阻抗谱研究了抗生物素蛋白/PAMAM/3-巯基丙酸(RSH)/金的性质、DNA的固定和杂交特性。序列特异性DNA的动态检测范围为1.4×10(-11)至2.7×10(-14) mol L(-1),检测限为1.4×10(-14) mol L(-1)。该DNA传感器不仅检测限低,而且对两碱基错配DNA表现出优异的选择性。

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