ULTRASENSITIVE METHODS FOR DNA QUANTITATION

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MPD仪器和专有的SuperTracer来增加探针的比活性。超级示踪剂以DNA探针为特征。采用多种符合方法来降低生物本底。初步实验证明，多重巧合探针方法允许相当大的拒绝非特异性生物背景。在PNA环结构中使用了非常特定的PNA试剂。总的来说，我们的研究证实，MPD允许改进qPCR方法和创新的直接（无目标扩增）核酸定量方法。这些创新方法的进一步发展是本研究的主要课题，特别是MPD增强型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配套试剂，能够在非常低的拷贝数下可靠地直接检测和定量DNA。这种检测方法将广泛应用于生物医学诊断活动，包括：临床诊断、治疗监测、药物发现和食品质量检测。我们将商业化一个由仪器和辅助试剂组成的检测系统，用于在非常低的拷贝数下进行DNA/RNA的定量检测。

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.