In situ hybridization chain reaction mediated ultrasensitive enzyme-free and conjugation-free electrochemcial genosensor for BRCA-1 gene in complex matrices
In situ hybridization chain reaction mediated ultrasensitive enzyme-free and conjugation-free electrochemcial genosensor for BRCA-1 gene in complex matrices
复制标题
原位杂交链式反应介导的超灵敏无酶、无缀合电化学基因传感器检测复杂基质中的 BRCA-1 基因
DOI：
10.1016/j.bios.2016.02.011
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发表时间：
2016-06-15
影响因子：
12.6
通讯作者：
Zhang, Guo-Jun
中科院分区：
文献类型：
--
作者：
Yang, Hui;Gao, Yang;Zhang, Guo-Jun
在这项研究中，我们报告了一种无酶和无连接的电化学基因传感器，能够超灵敏地读出乳腺癌易感基因BRCA-1。该传感器采用靶响应杂交链式反应(HCR)显著放大可检测到的电流信号。通过功能辅助探针对和通用的引发子序列，在靶序列存在的情况下，DNA寡聚体可以自发地和连续地聚合形成线性DNA串联结构。这种DNA纳米组装使基因传感器具有直到凌晨1点的超高灵敏度，比基于纳米材料或酶介导的扩增方法高出几个数量级。更重要的是，从凌晨1点到晚上10点，传感器的响应峰值电流与目标序列浓度的对数呈良好的线性相关此外，该传感器具有很高的选择性，可以区分单个不匹配的序列。这种基于HCR的基因传感器还能够直接在复杂的基质中探测低丰度的BRCA-1基因序列，例如50%的人血清，干扰最小。这些优势将使我们量身定制的基于HCR的电化学基因传感器在遗传分析和临床诊断方面具有吸引力。(C)2016爱思唯尔B.V.保留所有权利。
In this study, we report an enzyme-free and conjugation-free electrochemical genosensor enabling an ultrasensitive readout of BRCA-1, a breast cancer susceptibility gene. The sensor employs a target-responsive hybridization chain reaction (HCR) to significantly amplify the detectable current signals. By means of a functional auxiliary probe pair and a versatile initiator sequence, a linear DNA concatemer structure can be formed via spontaneous and continuous polymerization of DNA oligomers in the presence of target sequence. Such a DNA nanoassembly endows the genosensor an ultrahigh sensitivity up to 1 aM, which is higher than that of the nanomaterials-based or enzyme mediated amplification approaches by several orders of magnitude. More importantly, the sensor's responsive peak current exhibits a favorable linear correlation to the logarithm of the concentrations of target sequence ranging from 1 aM to 10 pM. In addition, the sensor is highly selective, and can discriminate a single mismatched sequence. This HCR-based genosensor is also capable of probing low-abundance BRCA-1 gene sequence directly in complex matrices, such as 50% human serum, with minimal interference. These advantages will make our tailor-engineered HCR-based electrochemical genosensor appealing to genetic analysis and clinical diagnostics. (C) 2016 Elsevier B.V. All rights reserved.