新型多目标物联合检测的量子点分子印迹功能化试纸制备及分析研究

The unsatisfactory detection efficiency and the unrealized joint detection are the two challenges in the anti-body free analysis application of molecular imprinting technology. In this project, test paper functionalized by molecular imprinted nanoparticles capped quantum dots would be prepared, and the detection method based on molecular imprinting combined with quantum dots would be build. By this method, three important biomarkers in prenatal screening would be detected simultaneously. In this project, macro-porous cellulose filter paper would be applied as the carrier for the molecular imprinted nanoparticles, to immediately separate the test paper from the sample solution. The nanoparticles acted as the solid ink could be printed on the surface of the filter paper, to improve the preparation efficiency of the test paper. Three kinds of imprinted nanoparticles with same excitation wavelength and different emission wavelength could be immobilized averagely on one piece of test paper, to realize the joint detection of the templates. This project would not only broaden the application field of the molecular imprinting technology, but also provide new technology and idea for the joint detection of prenatal screening with high performance. Furthermore, this project would promote the development of antibody-free analysis to solve the clinical problems.

针对目前分子印迹技术在无抗体分析领域应用过程中凸显出的检测效率低、检测对象单一等瓶颈问题，本项目拟通过制备集识别萃取、即时分离及多目标物联合检测的量子点分子印迹功能化试纸，发展基于量子点分子印迹技术的多目标物联合检测方法，并将其用于人血清样品中三种产前筛查标志物的联合检测。本项目创新之处在于：以大孔纤维素滤纸为量子点分子印迹纳米粒子载体，可实现印迹粒子从样品溶液中的即时分离，显著简化操作步骤；以量子点分子印迹纳米粒子为“固体墨水”打印在滤纸表面，可有效提高试纸制备效率；以激发波长相同、发射波长差异较大的三种量子点分子印迹纳米粒子为试纸表面检测单元，可实现对三种目标物的同时检测。本项目研究内容不仅能拓宽分子印迹技术应用范围、为产前筛查标志物的高效联合检测提供新技术与新思路，更有助于推动无抗体分析技术真正向解决临床实际问题方向发展。

分子印迹技术是通过模仿生命过程中抗原抗体特异性识别过程，采用人工合成方法，制备对模板分子具有高选择性识别能力的材料的技术，现已作为“人工抗体”在无抗体分析领域展现出良好的应用前景。本项目以进一步发展生命体内痕量标志物的无抗体分析技术为目标，成功设计并制备了四种基于荧光探针/量子点的分子印迹功能化材料。完成了相关表征及评价工作，开展了印迹材料对目标物的识别、吸附、检测及释放机理的探究工作，并在此基础上建立了相应的检测方法，实现了对复杂血清样品体系中甲胎蛋白等目标物的快速分析检测。本项目的实施，为生物标志物分离分析提供了准确性好、灵敏度高、稳定性好且成本低廉的检测技术，为推动无抗体分析技术向解决临床实际问题方向发展提供了技术支撑。