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Label-Free Telomerase Activity Detection via Electrochemical Impedance Spectroscopy

机译:通过电化学阻抗谱检测无标记端粒酶活性

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In the last decade, researchers have been searching for innovative platforms, methods, and techniques able to address recurring problems with the current cancer detection methods. Early disease detection, fast results, point-of-care sensing, and cost are among the most prevalent issues that need further exploration in this field. Herein, studies are focused on overcoming these problems by developing an electrochemical device able to detect telomerase as a cancer biomarker. Electrochemical platforms and techniques are more appealing for cancer detection, offering lower costs than the established cancer detection methods, high sensitivity inherent to the technique, rapid signal processing, and their capacity of being miniaturized. Therefore, Au interdigital electrodes and electrochemical impedance spectroscopy were used to detect telomerase activity in acute T cell leukemia. Different cancer cell concentrations were evaluated, and a detection limit of 1.9 × 105 cells/mL was obtained. X-ray photoelectron spectroscopy was used to characterize the telomerase substrate (TS) DNA probe self-assembled monolayer on gold electrode surfaces. Atomic force microscopy displayed three-dimensional images of the surface to establish a height difference of 9.0 nm between the bare electrode and TS-modified Au electrodes. The TS probe is rich in guanines, thus forming secondary structures known as G-quadruplex that can be triggered with a fluorescence probe. Confocal microscopy fluorescence images showed the formation of DNA G-quadruplex because of TS elongation by telomerase on the Au electrode surface. Moreover, electrodes exposed to telomerase containing 2′,3′-dideoxyguanosine-5′-triphosphate (ddGTP) did not exhibit high fluorescence, as ddGTP is a telomerase inhibitor, thus making this device suitable for telomerase inhibitors capacity studies. The electrochemical method and Au microchip device may be developed as a biosensor for a point-of-care medical device.
机译:在过去的十年中,研究人员一直在寻找能够解决当前癌症检测方法中反复出现的问题的创新平台,方法和技术。早期疾病检测,快速结果,即时护理感测和成本是最流行的问题,需要在该领域中进一步探索。本文中,研究集中在通过开发能够检测端粒酶作为癌症生物标记物的电化学装置来克服这些问题。电化学平台和技术对于癌症检测更具吸引力,与现有的癌症检测方法相比,其成本更低,该技术固有的高灵敏度,快速的信号处理以及其小型化能力。因此,使用Au叉指电极和电化学阻抗谱来检测急性T细胞白血病中的端粒酶活性。评价了不同的癌细胞浓度,检出限为1.9×105细胞/ mL。 X射线光电子能谱用于表征金电极表面上端粒酶底物(TS)DNA探针自组装单层。原子力显微镜显示表面的三维图像,以在裸电极和TS修饰的Au电极之间建立9.0 nm的高度差。 TS探针富含鸟嘌呤,因此形成了称为G-四链体的二级结构,可以用荧光探针触发。共聚焦显微镜荧光图像显示由于端粒酶在金电极表面上的TS延伸,导致DNA G四联体的形成。此外,由于ddGTP是端粒酶抑制剂,因此暴露于含有2',3'-二脱氧鸟苷-5'-三磷酸(ddGTP)的端粒酶的电极不显示高荧光,因此使该设备适合用于端粒酶抑制剂的研究。电化学方法和Au微芯片装置可以被开发为即时医疗装置的生物传感器。

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