基于核酸适配体-多功能胶体晶体编码微球液相芯片多元检测谷物中真菌毒素新体系研究

项目名称： 基于核酸适配体-多功能胶体晶体编码微球液相芯片多元检测谷物中真菌毒素新体系研究

项目编号： No.31471642

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 李建林

作者单位： 南京师范大学

项目金额： 86万元

中文摘要： 针对真菌毒素富集、净化、快速、低成本多元检测问题，通过纳米SiO2颗粒，Fe3O4颗粒共组装、凝胶及刻蚀技术等制备不同孔径及材料组成具有磁性和光学特性的多功能胶体晶体微球。以这些反射峰编码的微球为液相芯片载体，以谷物中典型真菌毒素为对象，研究微球表面修饰和核酸适配体（aptamer）的固定方法，研究应用该微球富集和净化谷物样品中真菌毒素的条件。设计真菌毒素aptamer和其互补链，构建基于aptamer作用原理的该微球真菌毒素液相芯片检测新方法。研究净化前后、aptamer互补链碱基长度、aptamer和互补链浓度比、杂交条件、不同孔径及材料对检测信号的影响，研究该检测方法的灵敏度，回收率，稳定性、重复性和特异性。在实际试样检测中和传统的高效液相色谱(HPLC)方法进行比较研究，建立基于aptamer-多功能胶体晶体微球编码载体富集、净化以及快速检测谷物中多元真菌毒素液相芯片新技术体系。

中文关键词： 核酸适配体；液相芯片；真菌毒素；多功能胶体晶体微球；多元检测

英文摘要： For enrichment, purification, fast, low-cost detection of multiplex mycotoxins, multifunctional colloidal crystal microspheres which had different pore size, material composition, magnetism and optical properties were fabricated by self-assembly of nano-SiO2, nano-Fe3O4, gel and chemical etching techniques. The surface modification and immobilization methods of mycotoxin aptamer on the microsphere surface were studied. The enrichment and purification conditions for the typical mycotoxins in grains were researched by the microspheres encoded by their reflection peak positions. Based on the specific reaction between the mycotoxin and its aptamer, the new multiplex detection system on the surface of multifunctional colloidal crystal microspheres was established by designing the mycotoxin aptamer and its anti-aptamer. The influence factors on detection signals were researched, which included enrichment, purification,concentrations of aptamer and anti-mycotoxin aptamer, oligonucleotide length of anti-mycotoxin aptamer, hybridization conditions, pore size and material composition of microspheres. The sensitivity, recovery ratio, stability, repeatability and specificity of the new detection system were studied. The application of the established assay method for multiplex mycotoxins in grains was performed and the results were compared with that of HPLC method. The new enrichment, purification and detection system for multiplex mycotoxins in grains was established by aptamer-multifunctional colloidal crystal microsphere suspension array.

英文关键词： aptamer;suspension array;mycotoxins;multifunctional colloidal crystal microsphere;multiplex detection