DNA nanostructure-based ultrasensitive electrochemical microRNA biosensor.
DNA nanostructure-based ultrasensitive electrochemical microRNA biosensor.
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DOI：
10.1016/j.ymeth.2013.07.035
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发表时间：
2013-12
期刊：
影响因子：
4.8
通讯作者：
Yanli Wen;Gang Liu;H. Pei;Lanying Li;Qin Xu;Wen Liang;Yan Li;Li Xu;Suzhen Ren;C. Fa
中科院分区：
文献类型：
--
作者：
Yanli Wen;Gang Liu;H. Pei;Lanying Li;Qin Xu;Wen Liang;Yan Li;Li Xu;Suzhen Ren;C. Fa
MicroRNA (miRNA) 是多种细胞过程的关键调节因子，由于其在血清中稳定存在，已被认为是有前途的癌症生物标志物。作为一种基于表面的电化学生物传感器，它为疾病相关 miRNA 的低成本即时护理测试 (POCT) 提供了巨大的机会。然而，miRNA 传感器的灵敏度常常受到传质和水电极界面的表面拥挤效应的限制。在这里，我们提出了使用基于 DNA 纳米结构的电化学 miRNA 生物传感器对 miRNA 进行超灵敏检测的协议和指南。通过采用基于三维DNA纳米结构的界面工程方法，我们可以直接检测到少至阿托摩尔（<1000拷贝）的miRNA，并且具有高单碱基辨别能力。由于这种超灵敏电化学 miRNA 传感器 (EMRS) 具有高度重复性，并且基本上无需事先进行靶标标记和 PCR 扩增，因此可以方便可靠地分析食管鳞状细胞癌 (ESCC) 患者临床样本中的 miRNA 表达水平。
MicroRNAs (miRNAs) are key regulators of a wide range of cellular processes, and have been identified as promising cancer biomarkers due to their stable presence in serum. As an surface-based electrochemical biosensors which offer great opportunities for low-cost, point-of-care tests (POCTs) of disease-associated miRNAs. Nevertheless, the sensitivity of miRNA sensors is often limited by mass transport and the surface crowding effect at the water-electrode interface. Here, we present a protocol as well as guidelines for ultrasensitive detection of miRNA with DNA nanostructure-based electrochemical miRNA biosensor. By employing the three-dimensional DNA nanostructure-based interfacial engineering approach, we can directly detect as few as attomolar (<1000 copies) miRNAs with high single-base discrimination ability. Since this ultrasensitive electrochemical miRNA sensor (EMRS) is highly reproducible and essentially free of prior target labeling and PCR amplification, it can conveniently and reliably analyze miRNA expression levels in clinical samples from esophageal squamous cell carcinoma (ESCC) patients.