纳米孔微流通道多元SERS传感技术用于病原菌基因诊断研究

项目名称： 纳米孔微流通道多元SERS传感技术用于病原菌基因诊断研究

项目编号： No.21275044

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 数理科学和化学

项目作者： 宦双燕

作者单位： 湖南大学

项目金额： 80万元

中文摘要： 本项目拟构建基于纳米孔的PDMS微流通道系统，集成纳米孔阻滞效应、生物传感信号放大技术和表面增强拉曼光谱（SERS）分析，发展高通量的食源性病原菌快速基因诊断新技术。利用生物分子识别事件将目标DNA阻滞在纳米孔前，结合等温扩增和酶循环切割等生物信号放大技术，定量剪切释放拉曼信号标记分子，采用纳米增强基底监测信号分子穿越纳米孔产生的SERS变化，发展高灵敏的食源性病原菌基因检测新技术，在构建单通道基础上设计三通道和辐射状多通道系统，发展高通量多元分析技术；结合错配或甲基化碱基识别蛋白和拉曼标记纳米粒子信号放大技术发展基因突变多元分析新方法；基于等位基因内切酶侵入剪切技术发展快速简便的"on-chip"高通量基因突变检测新技术；实现对不同致病菌种和突变位点的快速诊断和高灵敏检测。以期形成具有自主知识产权的关键方法和技术，建立食品安全监测和食源性病原菌快速检测的新技术平台。

中文关键词： DNA酶；DLISA；表面增强拉曼；细菌分析；多色荧光传感

英文摘要： This project is supposed to construct nanopore based PDMS microfluidic channels for multiplexed gene diagnosis of microbial pathogens in food,which combines the merits of resistive effect of nanopore with biosensing signal amplification techniques and sensitive Surface Enhanced Raman Scattering (SERS) analysis. Firstly, the microfluidic channels with nanopore will be constructed using PDMS matrix by rapid prototyping method.Then confocal raman microscope is coupled with the PDMS microfluidic system to establish a platform for biosensing. The recognition events of biomolecules captures the target DNA strand before the nanopore,then some signal amplification technologies, such as nicking enzyme assisted isothermal strand-displacement polymerization and enzymatic recycling cleavage for amplification, are utilized to quantitatively cleavage and release Raman signal labeling molecules. So the recognition events of pathogen gene sequence could be transformed into the detection of raman labeling molecules released. Surface enhanced nano-substrates would be used to monitoring the SERS signal chances from the raman tags that translocated through the nanopore to the detection areas. In this way, novel highly sensitive gene diagnosis methods would be constructed for multiplexed detection of microbial pathogens in food. Pro

英文关键词： DNAzyme；DLISA；SERS；Bacteria analysis；multiplexed ？uorescent analysis