Steichen M, Decrem Y, Godfroid E, Buess-Herman C
Service de Chimie Analytique et Chimie des Interfaces, Université Libre de Bruxelles, CP 255, Boulevard du Triomphe 2, B-1050 Bruxelles, Belgium.
Biosens Bioelectron. 2007 Apr 15;22(9-10):2237-43. doi: 10.1016/j.bios.2006.10.041. Epub 2006 Dec 12.
An electrochemical DNA hybridization detection method based on the electrostatic interactions of [Ru(NH3)6]3+ cations with the anionic phosphate backbone of DNA is proposed. PNA molecules are immobilized as capture probes on the gold substrate. The cationic ruthenium complexes do not interact electrostatically with the PNA probes due to the absence of the anionic phosphate groups on the PNA probes. But after hybridization, [Ru(NH3)6]3+ is adsorbed on the DNA backbone, giving a clear hybridization detection signal in ac voltammetry. The analytical parameters (sensitivity, selectivity and reproducibility) are evaluated. Very good discrimination against the single-base mismatch A2143G, internal to the 23S rRNA gene of Helicobacter pylori, is observed. Moreover the system is successfully applied to the detection of complementary PCR amplicons.
提出了一种基于[Ru(NH₃)₆]³⁺阳离子与DNA阴离子磷酸骨架静电相互作用的电化学DNA杂交检测方法。肽核酸(PNA)分子作为捕获探针固定在金基底上。由于PNA探针上不存在阴离子磷酸基团,阳离子钌配合物不会与PNA探针发生静电相互作用。但杂交后,[Ru(NH₃)₆]³⁺吸附在DNA骨架上,在交流伏安法中给出清晰的杂交检测信号。评估了分析参数(灵敏度、选择性和重现性)。观察到对幽门螺杆菌23S rRNA基因内部单碱基错配A2143G具有很好的区分能力。此外,该系统成功应用于互补PCR扩增子的检测。