Enzyme-free electrochemical biosensor based on amplification of proximity-dependent surface hybridization chain reaction for ultrasensitive mRNA detection
Enzyme-free electrochemical biosensor based on amplification of proximity-dependent surface hybridization chain reaction for ultrasensitive mRNA detection
复制标题
基于邻近依赖性表面杂交链式反应放大的无酶电化学生物传感器，用于超灵敏 mRNA 检测。
DOI：
10.1016/j.talanta.2020.121536
复制
发表时间：
2021-01-15
期刊：
影响因子：
6.1
通讯作者：
Jiang, Jian-Hui
中科院分区：
文献类型：
--
作者：
Cheng, Yu-Hong;Liu, Si-Jia;Jiang, Jian-Hui
在细胞中高效识别 mRNA 的能力将极大地促进 mRNA 介导的细胞级联及其疾病关联的阐明。然而，大多数传统的靶向核苷酸的电化学策略总是面临着繁琐的界面操作和洗涤程序，以及高昂的标记成本和严格的工具酶反应条件，限制了其潜在的应用。为了解决这些问题，我们在此首次报道了一种基于自主邻近依赖表面杂交链式反应（HCR）的简单无标记、等温、非酶促和超灵敏均相电化学生物传感器，用于灵敏信号放大和高度特异性检测目标生存素mRNA，检测限为3fM。该靶标在邻近依赖性表面杂交上触发二茂铁标记的亚稳态DNA发夹探针的杂交链式反应和mRNA驱动的表面杂交，从而形成多个长程双链DNA链，这些链通过基本稳定的二茂铁介导的氧化还原电流固定在金电极上。因此，根据目标 RNA 的浓度观察到显着的电化学信号增加，且检测限非常低。此外，该分子生物传感器还表现出优异的特异性，甚至可以区分单碱基错配，具有很强的可靠性。所开发的生物传感器为超灵敏和选择性检测提供了一种新型有前途的工具，并且在更详细地应用于mRNA相关生化研究和临床癌症诊断方面具有巨大的潜力。
The ability to recognize mRNA with high efficiency in cells would greatly facilitate the elucidation of mRNA-mediated cellular cascades and their disease associations. However, most traditional electrochemical strategies targeting nucleotides are always confronted with cumbersome interface operation and washing procedures, as well as the high cost of labeling and the strict reaction conditions of tool enzymes, limiting their potential applications. To address these issues, herein we reported, for the first time, a simple label-free, isothermal, non enzymatic, and ultrasensitive homogeneous electrochemical biosensor based on autonomous proximity dependent surface hybridization chain reaction (HCR), for sensitive signal amplification and highly specific detection of target survivin mRNA with a detection limit of 3 fM. The target triggers hybridization chain reaction and mRNA-fueled surface hybridization of ferrocene-tagged metastable DNA hairpin probes on proximity dependent surface hybridization, resulting in the formation of multiple long-range duplex DNA chains which are immobilized onto the gold electrodes with a substantially stable ferrocene-mediated redox current. Thus, a significant electrochemical signal increase is observed dependent on the concentration of the target RNA, with a very low detection limit. Mo-reover, this molecular biosensor also exhibits excellent specificity to distinguish even single base mismatched, with strong reliability. The developed biosensor provides a novel promising tool for ultra-sensitive and selective detection, and it has great potential to be applied in mRNA-related biochemical research and clinical cancer diagnostics in more detail.