新型探测禽流感病毒的糖荧光探针构建

The pandemic of influenza can cause a high death rate and severe economic loss worldwide (such as the newly discovered H7N9 in south China in 2013). However, conventional biochemical techniques for detection of influenza virus require costly facilities, onerous manipulations and long detection time, which limits their extensive use in practice. Development of simple, quick and accurate methods for the detection of the virus represents an urgent task to be addressed. This project takes advantage of the selective recognition between natural sialic acid-terminated trisaccharides and hemagglutinin on the surface of influenza virus particle for the design of a unique class of simple fluorescence probes for virus detection and discrimination. Easily prepared fluorescence glycodyes are used to couple with a quenching agent (either a small-molecule orgraphene) for the construction of fluorogenic virus probes. These probes will be used for the detection of various influenza viruses and for discrimination of their pandemic potential. Real samples from hospitals and poultry markets will be tested. This project aims at providing a reliable, quick and economic tool for the detection and estimation of influenza pandemic, thereby setting a basis for the further development of commercial agents for influenza diagnosis.

全球性禽流感大流行严重威胁人类生命，并造成社会经济严重损失。然而，现有对禽流感病毒的生化检测技术所使用设备昂贵，操作繁琐，耗时长，普适性差。发展快速高效的禽流感病毒检测新方法是国际上亟待解决的科学难题。本项目基于禽流感病毒颗粒表面血凝素与天然唾液酸三糖的高选择性结合能力，设计构建基于唾液酸三糖为识别基团，荧光染料为信号基团的简易糖荧光探针应用于病毒的快速测定。通过糖基与氨基荧光团成肟反应构建目标糖荧光分子，并利用分子间或分子与石墨烯间的堆叠效应搭建荧光“开启”型分子探针，实现对于纯化禽流感病毒株及突变病毒株（具备人传染特征）的特异检测与区分。进一步应用家禽市场和病毒感染者的临床样品验证所构建探针的实用价值。期待通过本项目的实施，为禽流感的快速测定及其大面积人传染的风险评估提供一类简易高效的普适性探测手段，为研发我国自主知识产权的病毒诊断试剂盒，替代现有国外垄断生化技术提供创新研究基础。

甲型流感病毒的季节性爆发严重威胁人类的生命健康，现有流感病毒检测手段主要依赖病毒培养分离法、血清学诊断法及病毒抗原检测法等，其存在成本高、处理周期长、灵敏度低等缺陷，对流感疫情突发情况下甲型流感病毒的快速、精准监测仍是当下生命健康领域的巨大挑战。本项目研究中，通过简易高效的点击化学反应和成肟反应合成了修饰唾液酸三糖基和五糖寡糖基的折叠型糖基荧光探针。利用唾液酸残基与甲型流感病毒颗粒表面糖识别血凝素发生选择性相互作用，实现了对不同亚型流感病毒的检测与区分。进一步合成了基于唾液酸糖基修饰的含不同荧光基团的寡糖荧光探针，利用具备强芳香吸附及荧光猝灭能力的二硫化钼材料作为载体，通过分子自组装技术构建了基于唾液酸三糖探针的荧光激活型糖簇探针，实现了临床来源人与禽流感病毒的协同检测。通过本项目的研究，发展了简便、灵敏、特异的甲型流感病毒检测新体系，为病毒诊断学的研究提供了新的化学工具。