液相生物芯片用功能微/纳米球的设计、制备及其在肝癌早期诊断上的应用

肝癌在我国死亡率仅次于肺癌和胃癌。该严重现状与临床上缺少灵敏、特异的早期诊断标志物和方法密切相关。本项目拟在前期研究基础上将多重宏量编码微球和聚集诱导发光（AIE）纳米球结合流式细胞仪构建蛋白和ctDNA甲基化标志物新型液相生物芯片多元检测双平台；利用红光锰掺杂硒化锌量子点的大Stokes位移和荧光寿命可调的特性，探索它和绿光量子点结合实现荧光微米球的四维宏量编码，并研究其在流式细胞仪上编码和解码的方法和途径；采用SPG膜乳化法制备AIE纳米球，利用其强聚集诱导荧光性能获得高检测灵敏度；选择临床常用的肝癌蛋白标志物和新发现的肝癌相关的ctDNA甲基化标志物为检测对象，探索并建立稳定性好、灵敏度高和线性范围宽的多指标检测技术平台; 经临床样本检测和ROC曲线分析评价检测性能和临床价值；为肝癌早期诊断提供高灵敏度、高特异性、快捷的多指标检测方法与技术平台，使更多肝癌患者受益于该技术。

Liver cancer has the third highest death rate among the cancers in China, which is closely related to limited effective biomarkers and diagnosis methods. This project attempts to combine doped quantum dots of Mn2+:ZnSe@ZnS encoded fluorescent microspheres and AIE nanobeads with the flow cytometry to build a novel high sensitive liquid biochip double-platform for proteins and ctDNA methylations. SPG membrane emulsification method will be used to achieve the controllable preparation of quantum dots encoded fluorescent microspheres and AIE nanobeads. After successfully achieving the ways of encoding and decoding, and investigating the effect of strong fluorescence of AIE nanobeads on detection sensitivity, the novel suspension liquid biochip platform will be used to realize high- throughput, fast joint detection of liver cancer specific protein and ctDNA methylation biomarkers, respectively, which will lay the foundation for further development of novel suspension liquid biochip with independent artificial intelligence, and provide an efficient new way for early diagnosis of liver cancer.