具有表面增强拉曼散射效应的复合纳米材料的制备及在病毒检测中的应用

自新型冠状病毒（COVID-19）大流行热点事件之后，公共卫生安全问题愈发成为各国关注的重点。控制传染源是应对类似传染性事件最有效的方法，这需要对传染源进行快速且高灵敏的识别，然而现有的病毒检测技术往往不能同时实现。本论文利用表面增强拉曼光谱（SERS）技术快速精准识别和高灵敏的特点，将SERS技术与现有的病毒检测技术相结合，制备了两种具有表面增强拉曼散射效应的复合纳米材料，并进行病毒检测的应用。主要研究内容和研究成果如下：

利用溶剂热法制备了粒径可控的空心结构Fe₃O₄纳米粒子（NPs），并实现了SiO₂壳层以及Au壳层的吸附与生长，通过对各组分的表征调控，最终成功制备出具有SERS效应的磁性Fe₃O₄@SiO₂@Au复合纳米材料。以严重急性呼吸系统综合征冠状病毒2型（SARS-CoV-2）的相关核酸片段作为目的片段，分别设计与目的核酸片段半互补的引物1以及引物2（修饰有拉曼报告分子CY5）。将引物1修饰在Fe₃O₄@SiO₂@Au NPs表面，并通过夹心法对目的核酸片段进行捕获，在粒子表面形成（引物1-目的片段-引物2）三明治结构。最后，利用拉曼光谱仪对Fe₃O₄@SiO₂@Au NPs进行检测，成功实现对SARS-CoV-2相关核酸片段浓度为10^-6 M的检出。

利用多元醇还原法制备了聚二烯丙基二甲基氯化铵（PDDA）修饰的单分散金八面体纳米粒子，并通过静电吸附作用围绕金八面体核心进行Au NPs以及拉曼报告分子吲哚菁绿（ICG）的组装。最后使用SiO₂壳层对Au/Au@ICG复合纳米结构进行封装，得到具有ICG分子拉曼信号的金复合纳米材料（Au/Au@ICG@SiO₂ NPs）。将Au/Au@ICG@SiO₂ NPs与侧流免疫层析技术结合，并利用拉曼光谱仪进行检测分析，实现对SARS-CoV-2的核衣壳蛋白500 fg/mL浓度的成功检出。相较于对侧流免疫层析试纸的传统生物识别方式4 pg/mL浓度的检测限，将对SARS-CoV-2核衣壳蛋白的检出能力提高了约1个数量级。这项研究展示了利用Au/Au@ICG@SiO₂ NPs结合侧流免疫层析技术对SARS-CoV-2进行高灵敏度检测的潜力，有望在类似传染性病毒的早期防控方面发挥重要作用。

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