本文關(guān)鍵詞： 腫瘤標(biāo)志物 光電傳感器 腫瘤細(xì)胞 納米探針　出處：《山東師范大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：癌癥是目前社會威脅人類發(fā)展的嚴(yán)重疾病,也是致死率很高的疾病之一,它的發(fā)病率每年呈上升的趨勢,因此,致力于癌癥的治療和預(yù)防的任務(wù)刻不容緩。癌癥的早期發(fā)現(xiàn)與診斷是預(yù)防癌癥的關(guān)鍵,對于癌癥,是否能早期診斷與發(fā)現(xiàn)并采取相應(yīng)的措施,直接關(guān)系到癌癥患者的治愈率和生存率。臨床現(xiàn)有的腫瘤檢測方法的特異性和靈敏度很低,對癌癥的早期發(fā)現(xiàn)仍存在很大困難,且確診較慢,容易耽誤最佳治療時(shí)機(jī)。為了提高臨床癌癥診斷方法的特異性和靈敏度,我們致力于低濃度腫瘤標(biāo)志物的檢測和腫瘤細(xì)胞的原位成像來實(shí)現(xiàn)癌癥的靈敏檢測。本論文針對腫瘤標(biāo)志物如甲胎蛋白(AFP)、micro RNA-21等,利用光電化學(xué)方法,實(shí)現(xiàn)了腫瘤標(biāo)志物和腫瘤細(xì)胞的原位分析。本論文主要包括以下內(nèi)容:1.基于石墨烯、鐵卟啉、金納米棒三元復(fù)合物開發(fā)了一種高效雙猝滅電致化學(xué)發(fā)光(ECL)免疫傳感器,通過具有類過氧化物酶性質(zhì)的三元復(fù)合物對H2O2催化作用和金納米棒(Au NRs)的共振能量轉(zhuǎn)移產(chǎn)生高強(qiáng)度的ECL信號。這種超靈敏ECL傳感策略,結(jié)合石墨烯/Au-Cd S量子點(diǎn)的優(yōu)良ECL傳感性能和雙猝滅反應(yīng),實(shí)現(xiàn)AFP的敏感檢測。2.首次合成新型石墨烯三重復(fù)合物石墨烯-鐵卟啉-金納米花(H-RGO-Au NFs),利用其優(yōu)良的類過氧化物酶的催化性能及適體技術(shù),設(shè)計(jì)一種新型電化學(xué)適體傳感器,可靈敏檢測白血病細(xì)胞(K562),并具有很好的選擇性。3.構(gòu)建信號放大型熒光納米探針,在金顆粒(Au NPs)上修飾DNA熒光探針,探針DNA可以結(jié)合細(xì)胞內(nèi)目標(biāo)miRNA,引發(fā)后續(xù)的DNA酶輔助的DNA鏈裂解,導(dǎo)致熒光基團(tuán)Cy5的釋放,Au NPs與被釋放熒光基團(tuán)Cy5間的較遠(yuǎn)的距離大大減弱了之前狀態(tài)的熒光猝滅效應(yīng),使Cy5產(chǎn)生熒光信號。此方法中一個目標(biāo)物miR-21和探針結(jié)合后可以產(chǎn)生一系列的剪切反應(yīng),進(jìn)而實(shí)現(xiàn)信號放大。因?yàn)閙i R-21在癌細(xì)胞中是過表達(dá)的,所以這種mi R-21的納米剪刀檢測為癌細(xì)胞的檢測提供了一種簡單高效的方法,并且可以應(yīng)用到細(xì)胞內(nèi)其他基因信號放大檢測和成像分析。
[Abstract]:Cancer is a serious disease threatening the development of human beings and one of the diseases with high mortality. The incidence of cancer is on the rise every year. The task of treating and preventing cancer is urgent. The early detection and diagnosis of cancer is the key to the prevention of cancer. It is directly related to the cure rate and survival rate of cancer patients. The specificity and sensitivity of the existing tumor detection methods are very low, the early detection of cancer is still very difficult, and the diagnosis of cancer is slow. In order to improve the specificity and sensitivity of clinical cancer diagnosis methods, We focus on the detection of low concentration tumor markers and in situ imaging of tumor cells to detect cancer sensitively. The in situ analysis of tumor markers and tumor cells has been achieved. In this thesis, the following contents are included: 1. Based on the ternary complexes of graphene, iron porphyrin and gold nanorods, an efficient double quenching electroluminescent chemiluminescence (ECL) immunosensor was developed. A high intensity ECL signal is generated by a ternary complex with peroxidase properties to catalyze H _ 2O _ 2 and the resonance energy transfer of gold nanorods au NRs. This super-sensitive ECL sensing strategy is developed. The excellent ECL sensing performance and double quenching reaction of graphene / Au-CdS quantum dots were studied. To realize the sensitive detection of AFP, a novel graphene triple complex, graphene, iron porphyrin-gold nano-flower, H-RGO-Au NFS was synthesized for the first time. A novel electrochemical aptamer sensor was designed by using its excellent peroxidase catalytic performance and aptamer technology. It can be used to sensitively detect K562G in leukemic cells, and has good selectivity. 3. To construct a large fluorescent nanoprobe with signal discharge and modify DNA fluorescence probe on au NPs of gold particles. The probe DNA could bind to the target miRNAs in the cell and induce the subsequent DNA chain cleavage assisted by DNA enzyme, resulting in the release of Cy5 from the fluorescent group Cy5 and the long distance between the released Cy5 group and the released fluorescence group Cy5, which greatly weakened the fluorescence quenching effect of the previous state. In this method, a target miR-21 and a probe can produce a series of shear reactions, which amplify the signal, because miR-21 is overexpressed in cancer cells. Therefore, the nanoscale scissors detection of miR-21 provides a simple and efficient method for the detection of cancer cells, and can be applied to other gene signal amplification detection and imaging analysis in cells.
【學(xué)位授予單位】：山東師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R730.4;TB383.1
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本文編號：1488605