microRNA-21与乙脑病毒新型光化学传感器的构建及其研究

发布时间：2018-04-11 17:32

本文选题：光化学传感器 + miRNA-21　；参考：《湘潭大学》2017年硕士论文

【摘要】：肿瘤标志物和病毒的检测对于疾病的诊断和治疗具有十分重要的意义,因此,发展简便快捷且灵敏度高的检测新方法势在必行。本文利用二氨基嘌呤(2-AP)独特的光学性质,构建了高灵敏检测肿瘤标志物mi RNA-21的荧光传感器;基于分子印迹技术和共振光检测技术构建了高特异性检测乙型脑炎病毒(JEV)的共振光传感器;主要研究工作有:1.设计了一种基于2-AP探针与信号放大技术的荧光传感器用于高灵敏、高选择性的检测mi RNA-21。该传感器利用G-四链体对2-AP荧光的保护和催化-发卡自组装(CHA)技术的信号放大作用提高灵敏度,检出限可低至4 p M。值得一提的是,该传感器不需要猝灭剂与酶的参与,简单、经济。此外,该传感器可以用来检测人类乳腺癌细胞裂解液中超常表达的mi RNA-21,表明该方法在癌症的早期诊断方面具有潜在的应用价值。2.提出了一种免猝灭剂、免酶的荧光传感器用于高灵敏度的检测mi RNA-21。传感器结合了2-AP独特的光学性质和CHA信号放大技术。与传统的荧光探针相比,2-AP探针不需要猝灭剂,这归因于2-AP独特的光学性质:在双链中的荧光强度低于在单链中的荧光强度。此外,信号放大技术免酶,使得传感器更加简单、经济。在最佳条件下定量检测目标mi RNA-21,得到了一个宽的线性范围:40 p M到40 n M(R2=0.999)。此外,提出的传感器还可以用来监测癌细胞中mi RNA-21的表达水平,结果表明该传感器在生物医学研究和早期临床诊断方面具有潜在的应用价值。3.设计了一种基于分子印迹的共振光传感器,用于快速,高选择性检测JEV。为了缩短吸附时间和减少非特异性吸附,我们采用表面分子印迹技术制备共振光传感器的识别元件:Fe_3O_4/Si O2磁性印迹粒子,制备的印迹粒子可以快速且高选择性的捕获目标病毒粒子,导致粒子尺寸增加,体系共振光增强。值得一提的是,所制备的印迹粒子能在20 min左右达到吸附平衡。此外,在选择性实验中,印迹粒子对目标病毒表现出了高特异性识别能力,并且非印迹粒子的非特异性吸附很低。最后通过血清样品加标实验证明该传感器在分离、检测病毒方面有一定的应用价值。
[Abstract]:The detection of tumor markers and viruses is of great significance for the diagnosis and treatment of diseases. Therefore, it is imperative to develop a simple, rapid and sensitive new detection method.Based on the unique optical properties of diaminopurine 2-APs, a highly sensitive fluorescent sensor for the detection of tumor marker mi RNA-21 was constructed.Based on molecular imprinting technique and resonance light detection technique, a high specific resonance light sensor for detecting Japanese encephalitis virus (JEV) was constructed.A fluorescence sensor based on 2-AP probe and signal amplification was designed to detect mi RNA-21 with high sensitivity and selectivity.The sensitivity of the sensor is improved by using the protection of G- quadruplex to 2-AP fluorescence and the signal amplification of catalytic-card self-assembly (char) technique. The detection limit can be as low as 4 p / M.It is worth mentioning that the sensor does not require quenching agents and enzyme participation, simple, economical.In addition, the sensor can be used to detect the abnormal expression of mi RNA-21 in human breast cancer cell lysate, indicating that this method has potential application value in early diagnosis of cancer.A kind of quenching agent, enzyme free fluorescence sensor, was proposed to detect mi RNA-21 with high sensitivity.The sensor combines the unique optical properties of 2-AP and the CHA signal amplification technology.Compared with the traditional fluorescence probe, 2-AP probe does not require quenching agent, which is due to the unique optical properties of 2-AP: the fluorescence intensity in double strand is lower than that in single strand.In addition, the signal amplification technology enzyme-free, making the sensor simpler and more economical.Under the optimum conditions, the target mi RNA-21 was quantitatively detected, and a wide linear range from: 40 pm to 40 n M ~ (2) R _ (2) (0.999) was obtained.In addition, the proposed sensor can also be used to monitor the expression of mi RNA-21 in cancer cells. The results show that the sensor has potential application value in biomedical research and early clinical diagnosis.A resonant light sensor based on molecular imprinting was designed for rapid and highly selective detection of JEV.In order to shorten the adsorption time and reduce the non-specific adsorption, we prepared the magnetic imprinted particles of the resonance light sensor by surface molecular imprinting technology:: Fe _ 3O _ 2 / Si _ 2O _ 2.The imprinted particles can capture the target virus particles quickly and selectively, resulting in the increase of particle size and the enhancement of resonance light.It is worth mentioning that the imprinted particles can reach adsorption equilibrium at about 20 min.In addition, in selective experiments, imprinted particles showed a high specificity for the target virus recognition, and the non-specific adsorption of non-imprinted particles was very low.Finally, it is proved that the sensor has certain application value in isolation and detection of virus.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3;TP212

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