基于RPA-NZs光电化学传感器的构建及其在恙虫病检测中的应用研究

Tsutsugamushi disease is a zoonotic disease caused by (Orientia tsutsugamushi, Ot) which seriously threatens human health. it is transmitted by the bite of trombiculid mite larvae (chiggers).The existing standard testing methods for tsutsugamushi disease have low sensitivity, complicated operation, time-consuming, and lack of effective methods on the early diagnosis of tsutsugamushi disease at present. In order to overcoming the bottleneck of traditional methods for detection of tsutsugamushi disease, the research adopt the advantage of technique for amplifying nucleic acid in vitro, new nano materials and photoelectrochemical bioanalysis. This program will develop a method for the detection of tsutsugamushi disease based on RPA, NZs and photoelectrochemical sensor, which promote the sensitivity of method via triple strategy based on signal amplification of nucleic acid detection through RPA, NZs loaded SA and a great number of HRPs and the high sensitivity of photoelectrochemical sensor. This program will integrate the advantages of molecular biology, nanotechnology and photoelectrochemistry and develop a new method for detecting nucleic acid, which not only provides a new direction for fast detecting tsutsugamushi disease, but also provides a new idea for detecting other biomolecules, and promote the development of interdisciplinary field of molecular biology, nanotechnology and photoelectrochemistry. The research has widely prospect in application.

恙虫病是由恙虫病东方体经恙螨幼虫叮咬传播的一种人兽共患传染病，严重威胁人体健康。现有恙虫病检测方法存在检测灵敏度低、操作复杂和检测费时等缺点，同时目前缺乏有效的恙虫病早期诊断方法。针对这些瓶颈问题，本项目联合体外核酸扩增技术、新型纳米材料和光电化学传感器三者优势，拟构建基于重组酶聚合酶扩增-纳米酶（RPA-NZs）的光电化学检测平台并用于恙虫病核酸早期快速检测。通过RPA体外核酸扩增技术放大核酸检测信号、利用NZs作为载体分子负载大量特异性分子链霉亲和素和辣根过氧化物酶提高检测灵敏度及光电化学高灵敏特性三方面策略提升恙虫病核酸检测灵敏度。本项目将分子生物学、纳米技术与光电化学有机结合，建立新的核酸检测平台，不仅为恙虫病的快速检测提供新方法，而且为其他生物分子检测提供新思路，同时促进相关交叉领域的发展，具有广泛的应用前景。

针对目前恙虫病检测方法存在检测灵敏度低、操作复杂和检测耗时等缺点，本项目通过联合重组聚合酶扩增技术（RPA）、TiO2纳米棒和光电化学传感器三者优势，构建了光电化学检测平台并用于恙虫病核酸早期快速检测。同时为了拓展多种光电纳米材料合成，开展了多种生物标志物的新型传感器和检测方法研究。主要包括：1）恙虫病核酸光电化学传感器的研究；2）免疫光电化学传感器的研究；3）MicroRNA检测新策略研究；4）光电纳米材料的合成及新型生物传感器的研究。本项目研究工在Bioelectrochemistry、ACS Applied Nano Materials和Biosensors and Bioelectronics等国际学术期刊上发表SCI论文9篇，申请国家发明专利4项。本项目研究成果在新型光电功能材料合成、生物传感器发展以及生物标志物检测和分析等方面具有学术和应用价值。