Benvidi Ali, Firouzabadi Afsaneh Dehghani, Moshtaghiun Seyed Mohammad, Mazloum-Ardakani Mohammad, Tezerjani Marzieh Dehghan
Department of Chemistry, Faculty of Science, Yazd University, Yazd, Iran.
Department of Chemistry, Faculty of Science, Yazd University, Yazd, Iran.
Anal Biochem. 2015 Sep 1;484:24-30. doi: 10.1016/j.ab.2015.05.009. Epub 2015 May 16.
We have designed a simple and novel electrochemical biosensor based on glassy carbon electrode (GCE) for DNA detection. GCE was modified with reduced graphene oxide (RGO) and gold nanoparticles (AuNPs) by the electrochemical method, which is helpful for immobilization of thiolated bioreceptors. The electrode modification processes were characterized by scanning electron microscopy (SEM) and electrochemical methods. Then a single-stranded DNA (ssDNA) probe for BRCA1 5382 insC mutation detection was immobilized on the modified electrode for a specific time. The experimental conditions, such as probe immobilization time and target DNA (complementary DNA) hybridization time and temperature with probe DNA, were optimized using electrochemical methods. The electrochemical response for DNA hybridization and synthesis was measured using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) methods. The calibration graph contains two linear ranges; the first part is in the range of 3.0×10(-20) to 1.0×10(-12)M, and the second segment part is in the range of 1.0×10(-12) to 1.0×10(-7)M. The biosensor showed excellent selectivity for the detection of the complementary sequences from noncomplementary sequences, so it can be used for detection of breast cancer.
我们设计了一种基于玻碳电极(GCE)的简单新颖的电化学生物传感器用于DNA检测。通过电化学方法用还原氧化石墨烯(RGO)和金纳米颗粒(AuNPs)对玻碳电极进行修饰,这有助于固定硫醇化生物受体。电极修饰过程通过扫描电子显微镜(SEM)和电化学方法进行表征。然后将用于检测BRCA1 5382 insC突变的单链DNA(ssDNA)探针在修饰电极上固定特定时间。使用电化学方法优化了诸如探针固定时间、目标DNA(互补DNA)与探针DNA的杂交时间和温度等实验条件。使用电化学阻抗谱(EIS)和循环伏安法(CV)测量DNA杂交与合成的电化学响应。校准曲线包含两个线性范围;第一部分在3.0×10(-20)至1.0×10(-12)M范围内,第二部分在1.0×10(-12)至1.0×10(-7)M范围内。该生物传感器对互补序列与非互补序列的检测显示出优异的选择性,因此可用于乳腺癌检测。