Department of Analytical Chemistry, Faculty of Pharmacy, Ege University, Izmir, Turkey.
Biosens Bioelectron. 2012 Jan 15;31(1):516-22. doi: 10.1016/j.bios.2011.11.029. Epub 2011 Nov 26.
An electrochemical genosensor for the detection of hypermethylation of the glutathione S-transferase P1 (GSTP1) gene, a specific marker of prostate cancer, was reported. This new sensor was used in combination with a single-use carbon graphite working electrode and differential pulse voltammetry, with the results of sample analysis based on the guanine oxidation signals obtained at +1.0 V before and after hybridization between probe and synthetic target or denatured PCR samples. The detected DNA hybridization was also characterized by electrochemical impedance spectroscopy with potassium ferri/ferrocyanide as a redox probe. The protocol consisted of 2 different modes: (i) capture probes selective for methylation-specific and unmethylated GSTP1 sequences were immobilized onto the sensor directly, and hybridization was formed on the electrode surface; (ii) probe/target or probe/noncomplementary target couples were mixed in solution phase, and the transducer was modified through simple adsorption. The limit of detection (S/N=3) was calculated as 2.92 pmol of target sequence in a 100-μl reaction volume. The optimum analytical detection parameters for the biosensor, as well as its future prospects, were also presented.
报道了一种电化学基因传感器,用于检测谷胱甘肽 S-转移酶 P1(GSTP1)基因的超甲基化,GSTP1 基因是前列腺癌的特异性标志物。该新型传感器与一次性使用的碳石墨工作电极和差分脉冲伏安法结合使用,根据杂交前后探针与合成靶标或变性 PCR 样品之间的杂交获得的+1.0 V 处鸟嘌呤氧化信号来分析样品。电化学阻抗光谱法也使用铁氰化钾/亚铁氰化钾作为氧化还原探针来表征检测到的 DNA 杂交。该方案包括 2 种不同的模式:(i)将针对甲基化特异性和非甲基化 GSTP1 序列的捕获探针直接固定在传感器上,并在电极表面形成杂交;(ii)探针/靶标或探针/非互补靶标对在溶液相中混合,然后通过简单吸附修饰换能器。在 100μl 反应体积中,目标序列的检测限(S/N=3)计算为 2.92pmol。还提出了生物传感器的最佳分析检测参数及其未来前景。