Dendrimers-based DNA biosensors for highly sensitive electrochemical detection of DNA hybridization using reporter probe DNA modified with Au nanoparticles
PAMAM dendrimer;
Au nanoparticles;
Self-assembly;
DNA biosensor;
QUARTZ-CRYSTAL MICROBALANCE;
PAMAM DENDRIMERS;
SENSOR;
ACID;
OLIGONUCLEOTIDE;
IMMOBILIZATION;
AMPLIFICATION;
MICROARRAYS;
TARGETS;
D O I:
10.1016/j.bios.2009.04.022
中图分类号:
Q6 [生物物理学];
学科分类号:
071011 ;
摘要:
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-NH2) to obtain a thin film. Single-stranded T-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(NH3)(6)](3+) 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. (C) 2009 Elsevier B.V. All rights reserved.