DNA Framework-Mediated Electrochemical Biosensing Platform for Amplification-Free MicroRNA Analysis.,Analytical Chemistry

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DNA Framework-Mediated Electrochemical Biosensing Platform for Amplification-Free MicroRNA Analysis.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-03-03 , DOI: 10.1021/acs.analchem.9b05616
Yanli Wen ₁ , Lanying Li ₁ , Jiang Li _{2,

3} , Meihua Lin ₄ , Gang Liu ₁ , Wen Liang ₁ , Li Xu ₁ , Yan Li ₁ , Xiaolei Zuo ₄ , Suzhen Ren ₁ , Ying Zhu _{2,

3}

Affiliation

MicroRNAs (miRNAs) have been explored as biomarkers for early diagnosis of diseases like cancers. However, it remains challenging to detect low-level miRNAs in the total RNA from real samples in a facile approach. In this work, we report a two-phase miRNA biosensing strategy based on a modular framework nucleic acid (FNA) platform, which combines the high efficiency of homogeneous reaction and the convenience of heterogeneous biosensing. In the first phase, free DNA probes bind target miRNAs in a homogeneous solution, forming a DNA-RNA complex with high base stacking energy. Then, at the second phase, the universal FNA interface on the electrode selectively mediated the transition of the complex from the solution onto the interface for electrochemical signal generating and transduction. By applying this method, we detected as few as 1 aM of miR-141, a cancer marker miRNA, without the need for nucleic acid amplification. The dynamic range spans 10 orders of magnitude. We demonstrate multiplex miRNA detection and discrimination of highly homologous miRNAs with mismatches as few as a single base. We also show that this system can detect miR-141 in only 50 ng of total RNA samples from real cells, which allows discrimination of prostate cancer cells with normal cells. We envision this platform may satisfy the need for facile and high-throughput screening of early cancer markers.

中文翻译：

DNA框架介导的电化学生物传感平台，可进行无扩增的MicroRNA分析。

微小RNA（miRNA）已被用作生物标志物，用于早期诊断癌症等疾病。然而，以简便的方法从真实样品中检测总RNA中的低水平miRNA仍然具有挑战性。在这项工作中，我们报告了基于模块化框架核酸（FNA）平台的两阶段miRNA生物传感策略，该策略结合了均相反应的高效率和异质生物传感的便利性。在第一阶段，游离的DNA探针在均质溶液中结合目标miRNA，形成具有高碱基堆积能量的DNA-RNA复合物。然后，在第二阶段，电极上的通用FNA界面选择性地介导了复合物从溶液到界面上的过渡，从而产生电化学信号并进行转导。通过应用此方法，我们检测到只有1 aM的miR-141（一种癌症标志物miRNA），而无需进行核酸扩增。动态范围跨越10个数量级。我们展示了多重miRNA的检测和高度同源的miRNA的鉴别，错配少至单个碱基。我们还显示，该系统仅能从真实细胞中检测到50 ng总RNA样品中的miR-141，从而可以区分正常细胞中的前列腺癌细胞。我们设想该平台可以满足对早期癌症标志物进行简便，高通量筛选的需求。我们还显示，该系统仅能从真实细胞中检测到50 ng总RNA样品中的miR-141，从而可以区分正常细胞中的前列腺癌细胞。我们设想该平台可以满足对早期癌症标志物进行简便，高通量筛选的需求。我们还显示，该系统仅能从真实细胞中检测到50 ng总RNA样品中的miR-141，从而可以区分正常细胞中的前列腺癌细胞。我们设想该平台可以满足对早期癌症标志物进行简便，高通量筛选的需求。

更新日期：2020-03-03

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