ULTRASENSITIVE METHODS FOR DNA QUANTITATION

DNA 定量的超灵敏方法

• 批准号: 6074590
• 负责人: Andrzej K Drukier
• 金额: $ 9.11万
• 依托单位: BIOTRACES, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6074590
• 关键词: biomedical equipment development; biopsy; method development; nucleic acid hybridization; nucleic acid probes; nucleic acid quantitation /detection; nucleic acids; peptides; polymerase chain reaction

We have successfully initiated the development of innovative methods for direct detection of nucleic acids. These methods rely on the use of supersensitive MPD instrumentation and proprietary SuperTracer to increase the specific activity of the probes. SuperTracers have been developed featuring a DNA probe. Multiple coincidence methods are being implemented to reduce biological background. Preliminary experiments documented that the multiple coincidence probe method permits a considerable rejection of non-specific biological background. Very specific PNA reagents in a PNA loop configuration were used. Overall, our studies confirmed that MPD permits improved qPCR methods and innovative direct (no target amplification) methods of nucleic acids quantitation. Further development of these innovative methods are main topics for the proposed research, especially the development of the MPD enhanced direct quantitation of DNA (dqDNA/MPD). PROPOSED COMMERCIAL APPLICATIONS: This project will develop procedures and supporting reagents which enable reliable direct detection and quantitation of DNA at very low copy number. Such an assay would have wide application in biomedical diagnostic activities, including: clinical diagnostics, therapy monitoring, drugs discovery and food quality testing. We will commercialize an assay system consisting of both instrumentation and supporting reagents to perform quantitative detection of DNA/RNA at very low copy number.

我们已经成功地启动了直接检测核酸的创新方法的开发。这些方法依赖于使用超灵敏的MPD仪器和专有的SuperTracer来增加探针的比活性。以DNA探针为特色的SuperTracer已经开发出来。正在实施多种符合方法来减少生物本底。初步实验证明，多重符合探针法允许对非特定生物背景有相当大的排斥作用。在PNA环构型中使用了非常特定的PNA试剂。总体而言，我们的研究证实，MPD允许改进的qPCR方法和创新的直接(无靶标扩增)核酸定量方法。进一步开发这些创新方法是拟议研究的主要主题，特别是开发MPD增强型DNA直接定量(dqDNA/MPD)。建议的商业应用：该项目将开发程序和支持试剂，使可靠的直接检测和定量的DNA在非常低的拷贝数。这种检测方法将在生物医学诊断活动中有广泛的应用，包括：临床诊断、治疗监测、药物发现和食品质量检测。我们将商业化一种由仪器和辅助试剂组成的检测系统，以极低的拷贝数进行DNA/RNA的定量检测。