Lee Jeong-Gun, Yun Kyusik, Lim Guei-Sam, Lee Sang Eun, Kim Suhyeon, Park Je-Kyun
Bio Electronics Group, LG Electronics Institute of Technology, 16 Woomyeon-dong, Seocho-gu, Seoul 137-724, South Korea.
Bioelectrochemistry. 2007 May;70(2):228-34. doi: 10.1016/j.bioelechem.2006.09.003. Epub 2006 Sep 23.
This paper reports a novel detection method for DNA hybridization based on the electrochemiluminescence (ECL) of Ru(bpy)(3)(2+) with a DNA-binding intercalator as a reductant of Ru(bpy)(3)(3+). Some ECL-inducible intercalators have been screened in this study using electrochemical methods combined with a chemiluminescent technique. The double-stranded DNA intercalated by doxorubicin, daunorubicin, or 4',6-diamidino-2-phenylindole (DAPI) shows a good ECL with Ru(bpy)(3)(2+) at +1.19 V (versus Ag/AgCl), while the non-intercalated single-stranded DNA does not. In order to stabilize the self-assembled DNA molecules during ECL reaction, we constructed the ECL DNA biosensor separating the ECL working electrode with an immobilized DNA probe. A gold electrode array on a plastic plate was assembled with a thru-hole array where oligonucleotide probes were immobilized in the side wall of thru-hole array. The fabricated ECL DNA biosensor was used to detect several pathogens using ECL technique. A good specificity of single point mutations for hepatitis disease was obtained by using the DAPI-intercalated Ru(bpy)(3)(2+) ECL.
本文报道了一种基于Ru(bpy)(3)(2+)的电化学发光（ECL）检测DNA杂交的新方法，该方法使用一种与DNA结合的嵌入剂作为Ru(bpy)(3)(3+)的还原剂。本研究采用电化学方法结合化学发光技术筛选了一些可诱导ECL的嵌入剂。由阿霉素、柔红霉素或4',6-二脒基-2-苯基吲哚（DAPI）嵌入的双链DNA在+1.19 V（相对于Ag/AgCl）时与Ru(bpy)(3)(2+)显示出良好的ECL，而非嵌入的单链DNA则没有。为了在ECL反应过程中稳定自组装的DNA分子，我们构建了一种ECL DNA生物传感器，用固定化的DNA探针将ECL工作电极隔开。在塑料板上的金电极阵列与一个通孔阵列组装在一起，寡核苷酸探针固定在通孔阵列的侧壁上。所制备的ECL DNA生物传感器用于通过ECL技术检测几种病原体。使用DAPI嵌入的Ru(bpy)(3)(2+) ECL获得了对肝炎疾病单点突变的良好特异性。
