ULTRASENSITIVE METHODS FOR DNA QUANTITATION
ULTRASENSITIVE METHODS FOR DNA QUANTITATION
批准号:
6074590
负责人:
Andrzej K Drukier
金额:
$9.11万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2000
资助国家:
美国
项目状态:
已结题
起止时间:
2000-02-01 至 2000-07-31
中文摘要
我们已成功开发了直接检测核酸的创新方法。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。SuperTracers已经开发出具有DNA探针的功能。正在实施多种符合方法以减少生物背景。 初步实验证明,多重符合探针方法允许相当大的拒绝非特异性生物背景。使用PNA环构型中的非常特异性的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(无靶扩增)核酸定量方法。这些创新方法的进一步发展是拟议研究的主要主题,特别是MPD增强的DNA直接定量(dqDNA/MPD)的发展。拟议的商业应用: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。这种分析将在生物医学诊断活动中具有广泛的应用,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支持试剂组成的测定系统,以在非常低的拷贝数下进行DNA/RNA的定量检测。
英文摘要
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.
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