發(fā)布時間：2018-04-11 17:32

  本文選題：光化學(xué)傳感器 + miRNA-21��； 參考：《湘潭大學(xué)》2017年碩士論文

【摘要】：腫瘤標(biāo)志物和病毒的檢測對于疾病的診斷和治療具有十分重要的意義,因此,發(fā)展簡便快捷且靈敏度高的檢測新方法勢在必行。本文利用二氨基嘌呤(2-AP)獨(dú)特的光學(xué)性質(zhì),構(gòu)建了高靈敏檢測腫瘤標(biāo)志物mi RNA-21的熒光傳感器;基于分子印跡技術(shù)和共振光檢測技術(shù)構(gòu)建了高特異性檢測乙型腦炎病毒(JEV)的共振光傳感器;主要研究工作有:1.設(shè)計了一種基于2-AP探針與信號放大技術(shù)的熒光傳感器用于高靈敏、高選擇性的檢測mi RNA-21。該傳感器利用G-四鏈體對2-AP熒光的保護(hù)和催化-發(fā)卡自組裝(CHA)技術(shù)的信號放大作用提高靈敏度,檢出限可低至4 p M。值得一提的是,該傳感器不需要猝滅劑與酶的參與,簡單、經(jīng)濟(jì)。此外,該傳感器可以用來檢測人類乳腺癌細(xì)胞裂解液中超常表達(dá)的mi RNA-21,表明該方法在癌癥的早期診斷方面具有潛在的應(yīng)用價值。2.提出了一種免猝滅劑、免酶的熒光傳感器用于高靈敏度的檢測mi RNA-21。傳感器結(jié)合了2-AP獨(dú)特的光學(xué)性質(zhì)和CHA信號放大技術(shù)。與傳統(tǒng)的熒光探針相比,2-AP探針不需要猝滅劑,這歸因于2-AP獨(dú)特的光學(xué)性質(zhì):在雙鏈中的熒光強(qiáng)度低于在單鏈中的熒光強(qiáng)度。此外,信號放大技術(shù)免酶,使得傳感器更加簡單、經(jīng)濟(jì)。在最佳條件下定量檢測目標(biāo)mi RNA-21,得到了一個寬的線性范圍:40 p M到40 n M(R2=0.999)。此外,提出的傳感器還可以用來監(jiān)測癌細(xì)胞中mi RNA-21的表達(dá)水平,結(jié)果表明該傳感器在生物醫(yī)學(xué)研究和早期臨床診斷方面具有潛在的應(yīng)用價值。3.設(shè)計了一種基于分子印跡的共振光傳感器,用于快速,高選擇性檢測JEV。為了縮短吸附時間和減少非特異性吸附,我們采用表面分子印跡技術(shù)制備共振光傳感器的識別元件:Fe_3O_4/Si O2磁性印跡粒子,制備的印跡粒子可以快速且高選擇性的捕獲目標(biāo)病毒粒子,導(dǎo)致粒子尺寸增加,體系共振光增強(qiáng)。值得一提的是,所制備的印跡粒子能在20 min左右達(dá)到吸附平衡。此外,在選擇性實(shí)驗(yàn)中,印跡粒子對目標(biāo)病毒表現(xiàn)出了高特異性識別能力,并且非印跡粒子的非特異性吸附很低。最后通過血清樣品加標(biāo)實(shí)驗(yàn)證明該傳感器在分離、檢測病毒方面有一定的應(yīng)用價值。
[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.
【學(xué)位授予單位】：湘潭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.3;TP212

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 徐群清;韋葉生;梁峰;梁麗艷;;IRMA法及ELISA法檢測HBV五項(xiàng)免疫學(xué)指標(biāo)的結(jié)果分析[J];中華實(shí)驗(yàn)和臨床病毒學(xué)雜志;2006年02期

相關(guān)碩士學(xué)位論文 前1條

1 周大學(xué);共振散射光譜法用于牛血清白蛋白的痕量檢測[D];重慶大學(xué);2012年

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本文編號：1736978