Electric-Field Assisted Isothermal On-Chip Amplification of DNA

电场辅助 DNA 片上等温扩增

• 批准号: 7052985
• 负责人: DALIBOR HODKO
• 金额: $ 15.54万
• 依托单位: NANOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-16 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7052985
• 关键词: biomedical equipment development; biotechnology; electric field; microarray technology; nucleic acid amplification techniques; portable biomedical equipment; temperature

DESCRIPTION (provided by applicant): Analysis of nucleic acids represents a well established molecular diagnostic test offering a highly specific detection of pathogens causing infectious diseases. Most clinical methods are based on the process of amplifying DNA and/or RNA molecules using polymerase chain reaction (PCR). The proposing team has recently developed another scheme for efficient isothermal amplification of DNA; the ramification amplification (RAM). This isothermal amplification is derived from primer extension, strand displacement, and the propagation of multiple ramification (branching) points. This technology utilizes a capture probe for target DNA or RNA isolation and a circularized RAM probe for subsequent amplification. Isothermal methods of DNA amplification have inherent advantages compared to the PCR method because there is no need to perform thermal cycling and can be easily packaged into a portable detection system. The proposed project combines two technologies, RAM amplification and electronic microarray technology where an electric field is used to assist and accelerate the DNA amplification process. The project is a joint effort in combining two compatible technologies and will leverage earlier efforts in RAM amplification and development of a portable DNA microarray detection system. The main advantage of the proposed approach is that a highly selective and sensitive RAM amplification will be integrated on the electronic microarray which will assure high level of multiplexing and rapid detection. Potential applications are envisioned in portable and/or point of care systems for detection of pathogens, infectious diseases and/or biological agents and emerging pathogens.

描述（由申请方提供）：核酸分析代表了一种成熟的分子诊断检测，可高度特异性地检测引起传染病的病原体。大多数临床方法基于使用聚合酶链式反应（PCR）扩增DNA和/或RNA分子的过程。该研究小组最近开发了另一种有效的DNA等温扩增方案：分枝扩增（RAM）。这种等温扩增源自引物延伸、链置换和多个分支（分支）点的繁殖。该技术利用捕获探针进行靶DNA或RNA分离，并利用环化RAM探针进行后续扩增。与PCR方法相比，DNA扩增的等温方法具有固有的优点，因为不需要进行热循环，并且可以容易地包装到便携式检测系统中。拟议的项目结合了两种技术，RAM扩增和电子微阵列技术，其中电场用于协助和加速DNA扩增过程。该项目是结合两种兼容技术的共同努力，并将利用早期在RAM扩增和便携式DNA微阵列检测系统开发方面的努力。所提出的方法的主要优点是，一个高度选择性和敏感的RAM扩增将集成在电子微阵列，这将确保高水平的多路复用和快速检测。在用于检测病原体、传染病和/或生物制剂和新兴病原体的便携式和/或护理点系统中设想潜在的应用。