基于SERS与DNA反应的高灵敏度检测和逻辑门传感

发布时间：2018-09-12 13:16

【摘要】：本文采用了核酸放大策略,构建了基于表面增强拉曼散射技术分别结合扩展分支杂交链式反应(EB-HCR)和滚环信号放大技术,实现了核酸放大以及miRNA高灵敏度检测和逻辑门传感,达到了预期的检测效果。主要研究包括如下两个方面:1.首先,开展了基于经典HCR和支化HCR设计的是扩展分支杂交链式反应(EB-HCR),目的是提供更多的分支;并通过EB-HCR和表面增强拉曼散射(SERS)的组合检测靶DNA实现高灵敏度和特异性。HCR扩展化,使得在自组装中的每一轮生长都发生更多的分枝,并结合SERS技术,信号大大增强,从而提高了灵敏度。还将转换步骤和信号扩增步骤纳入体系中,提高了靶DNA的低含量和不稳定性。通过信使DNA连接这三部分;信使DNA作为转换步骤的产物,还充当了扩展分支HCR扩增和SERS增强的接头。本方法具备较高的灵敏度,还证明了足够的特异性来区分具有相似序列的DNA。2.然后,在这项研究中,构建了一个逻辑系统,目的是鉴别癌症的组织起源,鉴别它们与mi RNA已建立的分布关系。该系统通过检测来自具有两个“门”的两组特征性miRNA(每个鉴定组输入三个miRNA,其中两种miRNA在两组中共用)。此系统以逻辑判断为中心,用多输入多输出逻辑门复合体实现;同时纳入SERS信号放大,为系统提供了处理mi RNA的实际能力和效率。证实了该系统成功解决了miRNA分布和癌症的组织起源多输入多输出逻辑关系;同时,还可以处理来自mi RNA的性质的其他问题,例如低浓度和不稳定性。
[Abstract]:In this paper, nucleic acid amplification strategy is used to construct the extended branched hybrid chain reaction (EB-HCR) and rolling ring signal amplification based on surface-enhanced Raman scattering, respectively. The amplification of nucleic acid, the high sensitivity detection of miRNA and the logic gate sensing are realized. The expected detection effect has been achieved. The main research includes the following two aspects: 1. Firstly, extended branched hybrid chain reaction (EB-HCR) based on classical HCR and branched HCR is designed to provide more branching, and high sensitivity and specificity. It makes each round of self-assembly grow more branched, and combined with SERS technology, the signal is greatly enhanced, thus improving the sensitivity. The low content and instability of target DNA were improved by incorporating the conversion step and signal amplification step into the system. The three parts are connected by messenger DNA, and Messenger DNA acts as the product of the conversion step and acts as the HCR amplification and SERS enhancement connector of the extended branch. The method has high sensitivity and sufficient specificity to distinguish DNA.2. with similar sequences. Then, in this study, a logical system was constructed to identify the tissue origin of cancer and their established distribution with mi RNA. The system detects two sets of characteristic miRNA from two "gates" (each identification group inputs three miRNA, and two kinds of miRNA are shared in the two groups). The system is based on logic judgment and is implemented with multi-input and multi-output logic gate complex, and the SERS signal is amplified, which provides the practical ability and efficiency for the system to deal with mi RNA. It is proved that the system can successfully solve the multiple input-multiple-output logic relationship between the distribution of miRNA and the tissue origin of cancer. At the same time, it can also deal with other problems from the properties of mi RNA, such as low concentration and instability.
【学位授予单位】：曲阜师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R440;O657.3

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