本文關(guān)鍵詞：基于核酸側(cè)向?qū)游龅膍icroRNA快速檢測(cè)新方法建立　出處：《合肥工業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 微小核糖核苷酸 金納米粒子 側(cè)向?qū)游黾夹g(shù) 表面增強(qiáng)拉曼光譜 多重檢測(cè)

【摘要】：隨著人們生活節(jié)奏的加快和飲食結(jié)構(gòu)的變化,罹患癌癥的人群增多,對(duì)癌癥的前期診斷和術(shù)后的監(jiān)控尤為重要。而MicroRNA的表達(dá)水平和癌癥的發(fā)生緊密相連。此外,microRNA可通過日常食物攝取進(jìn)入人體內(nèi),從而調(diào)控人體靶基因表達(dá)方式,進(jìn)而影響人體的生理功能。MicroRNA種類繁多,因此建立microRNA的快速、簡(jiǎn)單、高靈敏的多重檢測(cè)技術(shù)在醫(yī)學(xué)、食品營(yíng)養(yǎng)與安全等領(lǐng)域具有重要意義。本文建立了以下兩種檢測(cè)方法:(1)基于側(cè)向?qū)游龊怂嵩嚰垪l多重檢測(cè)microRNA本方法利用三組Au-SH probe-T probe“三明治”夾心結(jié)構(gòu)的形成實(shí)現(xiàn)對(duì)microRNA的三重檢測(cè)。在金納米粒子表面固定檢測(cè)探針SH probe,NC膜上固定捕獲探針T probe,目標(biāo)物microRNA可與SH probe和T Probe部分雜交,形成夾心結(jié)構(gòu)從而將金納米粒子捕獲并滯留在NC膜上,通過金納米粒子在檢測(cè)線上的累積從而出現(xiàn)肉眼可見的條帶。建立的單重核酸試紙條對(duì)microRNA-21,microRNA-155和microRNA-210肉眼檢測(cè)限分別為1 nM,5 nM和5 nM;建立的多重核酸試紙條可以很好地實(shí)現(xiàn)對(duì)三種目標(biāo)物的同時(shí)檢測(cè)且不發(fā)生交叉反應(yīng)。(2)基于側(cè)向?qū)游黾夹g(shù)的表面增強(qiáng)拉曼光譜雙重檢測(cè)microRNA本方法是建立在Au@Ag-SH probe-T probe夾心結(jié)構(gòu)形成的基礎(chǔ)上,在Au@Ag上固定檢測(cè)探針之前先在其表面修飾拉曼信號(hào)分子。當(dāng)修飾了拉曼信號(hào)分子的Au@Ag被滯留在檢測(cè)線上時(shí),通過肉眼根據(jù)檢測(cè)線的強(qiáng)度可以初步判斷待測(cè)物濃度,而通過拉曼光譜分析,在肉眼無法分辨的濃度范圍內(nèi)也能檢出信號(hào),從而提高檢測(cè)靈敏度,對(duì)microRNA-21和microRNA-155的拉曼檢測(cè)靈敏度可達(dá)到0.5 nM和1 nM。此外,根據(jù)Rho-6G和4-ATP拉曼特征峰的不同實(shí)現(xiàn)了在試紙條上單線檢測(cè)microRNA-21和microRNA-155。
[Abstract]:With the accelerated pace of life and changes in diet, cancer population increased early diagnosis and monitoring of cancer after surgery is particularly important. And the expression level of MicroRNA and cancer are closely linked. In addition, microRNA through the daily food intake into the body, the body to regulate the expression of target gene, and then physiological function of human.MicroRNA type is various, so the establishment of microRNA rapid, simple, high sensitive multi detection technology has important significance in medicine, nutrition and food security and other fields. This paper established the following two methods: (1) lateral chromatography nucleic acid strip multiple detection method using the microRNA Au-SH probe-T group formed three probe "sandwich" structure to achieve three detection based on microRNA. The gold nanoparticle surface fixed detection probe SH probe NC immobilized on the membrane The capture probe T probe, microRNA and SH can target probe and T Probe hybrid, which will form a sandwich structure of gold nanoparticles capture and retention in the NC membrane, the gold nanoparticles accumulated in the detection line to visible bands. A single nucleic acid test strip of microRNA-21, microRNA-155 and microRNA-210 eye detection limits were 1 nM, 5 nM and 5 nM; the establishment of the multiplex nucleic acid test strip can well realize three kinds of target detection at the same time and does not cross react. (2) surface enhanced Raman spectra of lateral tomography dual detection of microRNA this method is based on the formation of Au@Ag-SH probe-T probe sandwich structure based on Au@Ag before the fixed detection probe first coated on the surface of the Raman signal molecules. When modified Raman signal molecules of Au@Ag were stranded on the test line, through the eyes of flesh According to the detection of line intensity can determine the initial analyte concentration, and the Raman spectra analysis, signal was detected in the concentration range of the naked eye can not distinguish, thus improving the detection sensitivity of microRNA-21 and microRNA-155 Raman detection sensitivity can reach 0.5 nM and 1 nM. in addition, according to the Rho-6G and 4-ATP Raman peaks of different implementation in the test paper on the single detection of microRNA-21 and microRNA-155.

【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：Q78;O658.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 馮曉燕;夏成龍;陳瑩;王春鵬;儲(chǔ)富祥;;殼聚糖-銀納米粒子抗菌棉纖維滅菌機(jī)理初探[J];化工新型材料;2016年10期

2 黃巧蓮;金慶日;章先;周一媚;陳彥永;徐露凝;楊永春;程昌勇;田廣燕;桂海孌;方維煥;宋厚輝;;基于側(cè)向?qū)游鲈淼聂髑苟舅谹快速檢測(cè)試紙條的研制[J];浙江農(nóng)林大學(xué)學(xué)報(bào);2016年03期

3 王菡;李高華;張陽(yáng);陳濤;薛小平;;膠體金側(cè)向流免疫層析技術(shù)檢測(cè)有機(jī)磷農(nóng)藥殘留[J];食品安全質(zhì)量檢測(cè)學(xué)報(bào);2015年11期

4 李永玉;彭彥昆;孫云云;DHAKAL Sagar;郭浪花;;拉曼光譜技術(shù)檢測(cè)蘋果表面殘留的敵百蟲農(nóng)藥[J];食品安全質(zhì)量檢測(cè)學(xué)報(bào);2012年06期

5 ;米飯也能調(diào)控你的基因?[J];生物醫(yī)學(xué)工程與臨床;2011年06期

6 劉燕德;劉濤;孫旭東;歐陽(yáng)愛國(guó);郝勇;;拉曼光譜技術(shù)在食品質(zhì)量安全檢測(cè)中的應(yīng)用[J];光譜學(xué)與光譜分析;2010年11期

7 張燕;王強(qiáng)斌;;銀納米粒子的生物醫(yī)學(xué)應(yīng)用研究進(jìn)展[J];生物物理學(xué)報(bào);2010年08期

8 王靜;劉亞君;胡愛云;顧維閏;孫龍;;單分散的銀納米粒子的制備及表征[J];江蘇科技大學(xué)學(xué)報(bào)(自然科學(xué)版);2010年04期

9 景花;宋沁馨;周國(guó)華;;MicroRNA定量檢測(cè)方法的研究進(jìn)展[J];遺傳;2010年01期

10 王旭珍;翟培宇;曲江英;張海鷹;陳靜;;銀納米粒子的制備及其在光催化中的應(yīng)用[J];功能材料;2009年09期

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