Miti Andrea, Zuccheri Giampaolo
Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
Methods Mol Biol. 2018;1811:115-135. doi: 10.1007/978-1-4939-8582-1_8.
DNA biosensors could overcome some of the common drawbacks of lab-based techniques for nucleic acids detection for diagnostics purposes. One of the main impediments for such applications of DNA biosensors is their lack of sensitivity: this can prevent their full exploitation in the diagnostic analytical field. DNA nanotechnology could enhance DNA biosensors and let them perform at the required high sensitivity. Well-designed, programmable self-assembly reactions can be triggered by a specific nucleic acid target. The Hybridization Chain Reaction (HCR) is a self-assembly strategy in which the target nucleic acid sequence triggers the formation of long nicked double-stranded DNA nanostructures. This can be performed in solution or on a surface, and the process can be coupled to different signal transduction schemes. We here describe the methods to design and test HCR reactions for the detection of different nucleic acid targets in solution and the procedures to exploit this strategy on surfaces with an electrochemical biosensing platform.
DNA生物传感器可以克服基于实验室的核酸检测技术在诊断应用中的一些常见缺点。DNA生物传感器在这类应用中的主要障碍之一是其灵敏度不足:这可能会阻碍它们在诊断分析领域的充分应用。DNA纳米技术可以增强DNA生物传感器,并使其具有所需的高灵敏度。精心设计的可编程自组装反应可以由特定的核酸靶标触发。杂交链式反应(HCR)是一种自组装策略,其中靶核酸序列触发长切口双链DNA纳米结构的形成。这可以在溶液中或表面上进行,并且该过程可以与不同的信号转导方案相结合。我们在此描述了设计和测试用于检测溶液中不同核酸靶标的HCR反应的方法,以及在电化学生物传感平台上在表面利用该策略的程序。