【摘要】：本文采用了核酸放大策略,構(gòu)建了基于表面增強(qiáng)拉曼散射技術(shù)分別結(jié)合擴(kuò)展分支雜交鏈?zhǔn)椒磻?yīng)(EB-HCR)和滾環(huán)信號(hào)放大技術(shù),實(shí)現(xiàn)了核酸放大以及miRNA高靈敏度檢測(cè)和邏輯門傳感,達(dá)到了預(yù)期的檢測(cè)效果。主要研究包括如下兩個(gè)方面:1.首先,開展了基于經(jīng)典HCR和支化HCR設(shè)計(jì)的是擴(kuò)展分支雜交鏈?zhǔn)椒磻?yīng)(EB-HCR),目的是提供更多的分支;并通過EB-HCR和表面增強(qiáng)拉曼散射(SERS)的組合檢測(cè)靶DNA實(shí)現(xiàn)高靈敏度和特異性。HCR擴(kuò)展化,使得在自組裝中的每一輪生長(zhǎng)都發(fā)生更多的分枝,并結(jié)合SERS技術(shù),信號(hào)大大增強(qiáng),從而提高了靈敏度。還將轉(zhuǎn)換步驟和信號(hào)擴(kuò)增步驟納入體系中,提高了靶DNA的低含量和不穩(wěn)定性。通過信使DNA連接這三部分;信使DNA作為轉(zhuǎn)換步驟的產(chǎn)物,還充當(dāng)了擴(kuò)展分支HCR擴(kuò)增和SERS增強(qiáng)的接頭。本方法具備較高的靈敏度,還證明了足夠的特異性來(lái)區(qū)分具有相似序列的DNA。2.然后,在這項(xiàng)研究中,構(gòu)建了一個(gè)邏輯系統(tǒng),目的是鑒別癌癥的組織起源,鑒別它們與mi RNA已建立的分布關(guān)系。該系統(tǒng)通過檢測(cè)來(lái)自具有兩個(gè)“門”的兩組特征性miRNA(每個(gè)鑒定組輸入三個(gè)miRNA,其中兩種miRNA在兩組中共用)。此系統(tǒng)以邏輯判斷為中心,用多輸入多輸出邏輯門復(fù)合體實(shí)現(xiàn);同時(shí)納入SERS信號(hào)放大,為系統(tǒng)提供了處理mi RNA的實(shí)際能力和效率。證實(shí)了該系統(tǒng)成功解決了miRNA分布和癌癥的組織起源多輸入多輸出邏輯關(guān)系;同時(shí),還可以處理來(lái)自mi RNA的性質(zhì)的其他問題,例如低濃度和不穩(wěn)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.
【學(xué)位授予單位】：曲阜師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R440;O657.3

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