CASCADE ROLLING CIRCLE AMPLIFICATION FOR DNA DIAGNOSTICS

用于 DNA 诊断的级联滚环放大

• 批准号: 2792862
• 负责人: David C Thomas
• 金额: $ 42.94万
• 依托单位: CANCER DETECTION TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2000-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2792862
• 关键词: DNA primers; diagnosis design /evaluation; fluorescence resonance energy transfer; genetic screening; genetic techniques; molecular cloning; technology /technique development

DESCRIPTION (Applicant's abstract): A novel isothermal DNA amplification technique has been developed called cascade rolling circle amplification (CRCA). This method is simple, rapid, and highly sensitive, achieving over a billion-fold amplification within one hour. By making one of the primers an energy transfer-labeled primer that generates a fluorescent signal only when incorporated into the amplicon, CRCA products can be detected within a closed system. These features make this technology suitable for a variety of diagnostic applications including both automated platforms and point-of-care field devices. Phase I research has demonstrated the feasibility of CRCA for DNA diagnostics, including detection of pathogenic agents. The objectives of this Phase II project include i) improvements in the assay to make the technology more adaptable to a variety of platforms, ii) development of multiplex assay using energy transfer-labeled primers of different colors, iii) optimizing the system for real-time detection to accurately quantitate target sequences, iv) adapting CRCA to a point-of-care diagnostic platform that is non-fluorescence based, and v) adapting the technology to an array platform. Long-range objectives include implementing CRCA in an automated amplification/detection system and developing broad applications for the point-of-care diagnostic market PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述(申请人摘要)：一种新的等温DNA扩增 已经开发出一种称为串级滚圆放大的技术 (CRCA)。该方法简单、快速、灵敏度高，达到了 在一小时内放大数十亿倍。通过使其中一条引子成为 能量转移标记的底漆，仅在以下情况下才产生荧光信号 纳入扩增子，CRCA产品可在闭合范围内检测 系统。这些特性使该技术适用于各种 诊断应用程序，包括自动化平台和医疗点 现场设备。第一阶段研究证明了CRCA在以下方面的可行性 DNA诊断，包括病原体检测。的目标 这个第二阶段的项目包括：一)改进化验方法，使 技术更能适应各种平台，ii)开发 用不同颜色的能量转移标记引物进行多重分析，III) 优化实时检测系统，准确量化目标 序列，iv)使CRCA适应于护理点诊断平台，即 以及v)使该技术适用于阵列平台。 长期目标包括在自动化环境中实施CRCA 扩增/检测系统和开发广泛的应用 医疗点诊断市场 建议的商业应用：不可用