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).
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.
