基于万古霉素核酸多聚体和核酸保护放大的可控纳米组装生物传感器的构建及致病菌检测

To date, biosensors for pathogenic bacteria detection have some drawbacks towards versatility, stability, sensitivity and signal outputs. In order to address the issues above mentioned, based on our previous works, we plan to design vancomycin(Van) modified single DNA strains to construct two sets of Van-DNA polymers in one-way or two-ways stretch. This creative design overcomes the shortcomings of reported vancomycin polymers, providing a optimized recognization component with‘broad spectrum’towards gram-positive pathogenic bacterial strains. Moreover, due to the smart design of Van-DNA polymers, the techniques of terminal protection of small-molecular modified DNA and rolling circling amplification have been introduced, transferring bacterial recognizing signal to circling amplified DNA signal, which could organize the assembly of functional nanoparticles and lead to SPR and SERS signals with high sensitivity and good stability. This biosensor of controllable assembly of nanoparticles based on Van-DNA polymer and DNA protection/amplification techniques, could detect gram-positive bacterial strains with good versatility, high sensitivity, strong stability and multi-signal output, which is very significant towards food safety and enviromental monitoring.

目前，用于致病菌检测的生物传感器在通用性、稳定性、灵敏度以及信号输出方面存在诸多问题。为解决这些问题，申请人在前期工作基础上，拟采用万古霉素偶联的单链核酸分子，通过碱基互补配对，巧妙地设计构建单向延伸和双向延伸两种模式的万古霉素核酸(Van-DNA)多聚体系列。一方面，这一创新性设计弥补了现有万古霉素多聚体存在的缺陷，为所有革兰氏阳性致病菌提供了一个全面优化的“广谱”识别元件。另一方面，通过Van-DNA多聚体系列的巧妙设计，还可以引入核酸末端保护和滚环扩增技术，将致病菌的识别信号转化为循环放大的核酸信号，并以扩增放大的核酸分子为基质介导功能化纳米颗粒的有序组装，产生超高灵敏度和良好稳定性的SPR和SERS多种传感信号。这一基于Van-DNA多聚体和核酸保护放大技术的可控纳米组装生物传感器能够实现革兰氏阳性致病菌的“广谱”、超灵敏、高稳定性的多信号检测，对食品安全、环境监测等具有重要意义。

用于致病菌检测的生物传感器在通用性、稳定性、灵敏度以及信号输出方面存在诸多问题。为解决这些问题，本课题以万古霉素偶联的单链核酸分子为基础，借助碱基互补配对，设计构建了单向延伸模式的万古霉素核酸(Van-DNA)二聚体。该二聚体作为靶向分子，具有成本低﹑生物相容性好并且构象可控等优点。另外，通过Van-DNA二聚体系列的巧妙设计，引入了核酸末端保护和滚环扩增技术，将致病菌的识别信号转化为循环放大的核酸信号，并以扩增放大的核酸分子为基质介导功能化纳米颗粒的有序组装，产生超高灵敏度和良好稳定性的SPR和SERS多种传感信号。这一基于Van-DNA二聚体和核酸保护放大技术的可控纳米组装生物传感器能够实现革兰氏阳性致病菌的“广谱”、超灵敏、高稳定性的多信号检测，对食品安全、环境监测等具有重要意义。围绕本课题，在国际专业期刊杂志《Colloids and Surfaces B: Biointerfaces》上发表论文2篇，《Nano》和《高等学校化学学报》等其他期刊上各发表1篇，总计发表标注基金资助的SCI论文6篇。同时申请国家发明专利3项，其中已经授权2项。基于本课题，现已培养硕士研究生3名。