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利用滚环扩增提高硅纳米线生物传感器的信噪比。

Signal-to-noise ratio enhancement of silicon nanowires biosensor with rolling circle amplification.

机构信息

State Key Laboratories of Transducer Technology & Science and Technology on Micro-system Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences , Shanghai, 200050, China.

出版信息

Nano Lett. 2013 Sep 11;13(9):4123-30. doi: 10.1021/nl401628y. Epub 2013 Aug 15.

Abstract

Herein, we describe a novel approach for rapid, label-free and specific DNA detection by applying rolling circle amplification (RCA) based on silicon nanowire field-effect transistor (SiNW-FET) for the first time. Highly responsive SiNWs were fabricated with a complementary metal oxide semiconductor (CMOS) compatible anisotropic self-stop etching technique which eliminated the need for hybrid method. The probe DNA was immobilized on the surface of SiNW, followed by sandwich hybridization with the perfectly matched target DNA and RCA primer that acted as a primer to hybridize the RCA template. The RCA reaction created a long single-stranded DNA (ssDNA) product and thus enhanced the electronic responses of SiNW significantly. The signal-to-noise ratio (SNR) as a figure-of-merit was analyzed to estimate the signal enhancement and possible detection limit. The nanosensor showed highly sensitive concentration-dependent conductance change in response to specific target DNA sequences. Because of the binding of an abundance of repeated sequences of RCA products, the SNR of >20 for 1 fM DNA detection was achieved, implying a detection floor of 50 aM. This RCA-based SiNW biosensor also discriminated perfectly matched target DNA from one-base mismatched DNA with high selectivity due to the substantially reduced nonspecific binding onto the SiNW surface through RCA. The combination of SiNW FET sensor with RCA will increase diagnostic capacity and the ability of laboratories to detect unexpected viruses, making it a potential tool for early diagnosis of gene-related diseases.

摘要

本文首次应用滚环扩增(RCA)技术,结合硅纳米线场效应晶体管(SiNW-FET),描述了一种快速、无标记、特异性的 DNA 检测新方法。采用与互补金属氧化物半导体(CMOS)兼容的各向异性自停止刻蚀技术制备高响应性 SiNW,无需混合方法。探针 DNA 固定在 SiNW 表面,然后与完全匹配的靶 DNA 和 RCA 引物进行三明治杂交,RCA 引物作为引物与 RCA 模板杂交。RCA 反应产生长的单链 DNA(ssDNA)产物,从而显著增强 SiNW 的电子响应。信号噪声比(SNR)作为一种衡量标准,用于分析信号增强和可能的检测极限。纳米传感器显示出对特定靶 DNA 序列的高灵敏度、浓度依赖性电导变化。由于 RCA 产物的大量重复序列的结合,实现了 >20 的 SNR,用于检测 1 fM DNA,这意味着检测下限为 50 aM。由于 RCA 大大减少了非特异性结合到 SiNW 表面的数量,因此基于 RCA 的 SiNW 生物传感器还能够从单碱基错配 DNA 中完美区分出完全匹配的靶 DNA,具有很高的选择性。SiNW FET 传感器与 RCA 的结合将提高诊断能力和实验室检测意外病毒的能力,使其成为基因相关疾病早期诊断的潜在工具。

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