Xu Lijian, Du Jingjing, He Nongyue, Deng Yan, Li Song, Wang Ting
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, P. R. China.
J Nanosci Nanotechnol. 2009 Apr;9(4):2698-703. doi: 10.1166/jnn.2009.dk09.
A novel and sensitive electrochemical DNA biosensor based on porous pseudo-carbon paste electrode (PPCPE) for DNA immobilization and enhanced hybridization detection is described. PPCPE was fabricated by mixing polymethyl methacrylate (PMMA) microspheres as the template, graphite powders as the filler, pyrrole as the precursor of polymer which actually acted as the paste. After the polymerization of pyrrole catalyzed by Fe3+, the template PMMA microspheres were removed to form PPCPE. The pore size were determined by SEM observations, their diameters were in the range from 2 to 5 microm, and the specific surface area of PPCPE was 42.76 m2/g by N2 adsorption at 77 K using the Brunauer Emmet Teller (BET) method. The hybridization reaction on the electrode was monitored by anodic stripping voltammetry (ASV) analysis using the method of gold nanoparticle-catalyzed silver enhancement. Compared with previous DNA sensors the attached DNA and complementary DNA detection sensitivity were dramatically increased. The experiments of ASV showed that the peak current of Ag at PPCPE were linear with the amount of complementary oligonucleotide ranging from 1 to 80 nM and 0.08 to 1 nM, the detection limit was as low as 0.05 nM.
描述了一种基于多孔伪碳糊电极(PPCPE)的新型灵敏电化学DNA生物传感器,用于DNA固定和增强杂交检测。PPCPE通过将聚甲基丙烯酸甲酯(PMMA)微球作为模板、石墨粉作为填料、吡咯作为实际上充当糊剂的聚合物前体混合制备而成。在Fe3+催化吡咯聚合后,去除模板PMMA微球以形成PPCPE。通过扫描电子显微镜(SEM)观察确定孔径,其直径范围为2至5微米,使用布鲁诺尔-埃米特-特勒(BET)方法在77 K下通过N2吸附测得PPCPE的比表面积为42.76 m2/g。使用金纳米颗粒催化银增强法通过阳极溶出伏安法(ASV)分析监测电极上的杂交反应。与先前的DNA传感器相比,附着DNA和互补DNA的检测灵敏度显著提高。ASV实验表明,PPCPE上Ag的峰值电流与互补寡核苷酸的量在1至80 nM和0.08至1 nM范围内呈线性关系,检测限低至0.05 nM。