基于上转换荧光纳米粒子表面核酸等温延伸microRNA信息检测技术研究

项目名称： 基于上转换荧光纳米粒子表面核酸等温延伸microRNA信息检测技术研究

项目编号： No.61301039

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 无线电电子学、电信技术

项目作者： 张立明

作者单位： 湖南工业大学

项目金额： 28万元

中文摘要： 研究表明microRNA(miRNA)在癌症、心血管等重大疾病中表达异常，获得这些特异性miRNA表达信息可为疾病早期诊断提供重要依据。但由于miRNA自身特点，其检测较难。目前检测方法存在特异性不高、费用昂贵、操作复杂等缺点。本项目结合碱基堆积原理、等温延伸技术以及荧光共振能量转移（FRET）原理，建立新型的miRNA信息获取平台。该平台利用目标miRNA提供的碱基堆积力帮助特异性引物与连接在NaYF4:Yb,Er荧光纳米粒子表面的通用引物互补配对，并进行等温延伸。在延伸的过程中，引入Cy3标记的dUTP。从而以荧光纳米粒子为能量供体，Cy3为受体，构建FRET体系。无目标miRNA存在时，在操作温度下特异引物与通用引物不能结合，因而不能发生延伸。本方法无需反转，操作简单，且通过FRET可有效降低背景干扰达到高灵敏检测的目的，适合早期诊断与临床检测。

中文关键词： 上转换荧光纳米颗粒；miRNA；热分解法；内参；牛血清白蛋白

英文摘要： Increasing evidence indicates that aberrant microRNA(miRNA) expression is a common feature of human diseases including cancers, cardiovascular disorders, metabolic disturbances and other serious diseases. These biomolecules have great potential for the early diagnosis of human diseases. However, several unique characteristics of miRNAs, including their small size, sequence homology among family members, and low abundance in total RNA samples, make it difficult to analyze the miRNA. Accordingly, the current technologies to detect miRNA have many drawbacks, such as low sensitivity and specificity, expensivity, and time-exaustivity. Herein, we propose a high sensitive and specific, and simple-executive miRNA imformation obtaining technique based on isothermal extension of nuclear acids chain on the surface of NaYF4:Yb,Er upconversion nanoparticles. An exponential extension process is only happened efficiently in the presence of target miRNA which provides additional stability for the universal primer on the surface of upconversion nanoparticles and specific primer in the solution. Associated by the DNA polymerase, both universal primer and specific primer will be extented and Cy3-labeled dUTP will be inserted. Thus, an effective fluorescence resonance energy transfer(FRET) system will be constructed. In this FRET

英文关键词： Upconversion fluorescence nanoparticles；miRNA；Thermal decomposition method；Reference；Bovine serum albumin