三种人畜共患传染性病原菌同步快速富集与检测技术研究

项目名称： 三种人畜共患传染性病原菌同步快速富集与检测技术研究

项目编号： No.81473018

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 李娟

作者单位： 吉林大学

项目金额： 80万元

中文摘要： 近年来，人畜共患传染病已成为新发公共卫生传染病的主要疾病，加强对其同步、快速、准确的检测技术研究十分必要。本课题是利用融合蛋白技术设计合成布鲁氏菌、大肠杆菌O157:H7以及单增李斯特菌三种发病率较高的人畜共患病传染性病原菌的特异融合蛋白抗原，联合磁纳米技术和量子点标记技术，利用免疫学的原理实现对待检样品中三种病原菌同步富集与快速准确检测。课题的主要研究内容包括：①表面功能化磁性纳米粒子的制备及表征；②三种病原菌特异融合蛋白抗原及其鸡卵黄抗体IgY的制备及鉴定；③磁性纳米生物探针的制备；④三种病原菌兔多克隆抗体及免疫化荧光量子点的制备；⑤借助荧光显微成像技术和荧光光谱仪对待检样品进行同步、快速定性及定量检测。课题研究的目的是设计制备具有我国自主知识产权和广泛应用前景的能同步富集三种病原菌的磁性纳米生物探针和荧光量子点生物探针，实现对三种常见人畜共患传染性病原菌同步快速、准确检测。

中文关键词： 人畜共患传染病；布鲁氏菌；大肠杆菌O157；H7；单核细胞增生性李斯特菌；检测技术

英文摘要： In recent years,zoonosis has become a main emerging public infectious disease .It is necessary to study on the synchronous,rapid and accurate detection technologies of its ralated pathogens. This topic is to design a specific fusion protein antigen of three kinds of zoonotic infectious pathogens with higher prevalence including Brucella, escherichia coli O157: H7 and L.monocytogenes by using of fusion protein technology,and to obtain a method aiming at the synchronization enrichment and rapid-accurate detection of these three pathogens in samples by using magnetic nanotechnology and quantum dot labeling technique and the principles of immunology. This topic research contents include:① preparation and characterization of surface-functionaled magnetic nanoparticles;② preparation and characterization of three kinds of pathogenic bacteria specific antigen fusion protein and their yolk antibody（IgY）; ③ the preparation of magnetic nano-biological probe; ④ preparation of rabbit polyclonal antibodies of three kinds of pathogenic bacterias and their immunization fluorescence quantum dots; ⑤ Synchronous,rapid,qualitative and quantitative analysis of the three kinds of antigens in samples by means of fluorescence microscopic imaging technology and fluorescence spectrometer.The purpose of this research is to design and prepare magnetic nano-biological probe and fluorescent quantum dot bio-probes with our own intellectual property and broad prospect ,which can be used to enrich these three common zoonotic infectious pathogens synchronously and detect them rapidly and accurately.

英文关键词： Zoonosis;Brucella;E.coli O157:H7;Listeria monocytogenes;Detection technology