Ilkhani Hoda, Farhad Siamak
Advanced Energy & Sensor Lab., Department of Mechanical Engineering, The University of Akron, Akron, OH 44325, United States.
Anal Biochem. 2018 Sep 15;557:151-155. doi: 10.1016/j.ab.2018.06.010. Epub 2018 Jun 14.
The aim of this study was to fabricate a novel electrochemical-based DNA-sensing device for Ebola virus DNA diagnostic by an enzyme-amplified detection, which improves the sensitivity and selectivity of the sensor. A thiolated DNA capture probe sequence was immobilized on the screen printed electrode surface and hybridized with biotinylated target strand DNA for the fabrication of Ebola DNA-sensing devices. Prior to the electrochemical detection of the enzymatic product by differential pulse voltammetry (DPV) method, the biotinylated hybrid was labeled with a streptavidin-alkaline phosphatase conjugate on the surface of the working electrode. All the experiment steps were optimized using electrochemical impedance spectroscopy (EIS) and the optimum condition for biosensor fabrication was achieved. A detection limit of 4.7 nM complementary oligonucleotides was obtained using the fabricated biosensor and the standard deviation of the blank solution. The last step, the selectivity and reproducibility of fabricated electrochemical DNA biosensor was obtained.
本研究的目的是通过酶放大检测制造一种新型的基于电化学的用于埃博拉病毒DNA诊断的DNA传感装置,以提高传感器的灵敏度和选择性。将硫醇化DNA捕获探针序列固定在丝网印刷电极表面,并与生物素化的靶链DNA杂交,以制造埃博拉DNA传感装置。在通过差分脉冲伏安法(DPV)对酶产物进行电化学检测之前,将生物素化的杂交体用链霉亲和素-碱性磷酸酶共轭物标记在工作电极表面。使用电化学阻抗谱(EIS)对所有实验步骤进行了优化,并实现了生物传感器制造的最佳条件。使用制造的生物传感器和空白溶液的标准偏差获得了4.7 nM互补寡核苷酸的检测限。最后一步,获得了制造的电化学DNA生物传感器的选择性和重现性。