METHODS FOR DETERMINING THE LENGTH OF TRIPLET REPEATS

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We are developing innovative methods for the direct, i.e. non-amplified, detection of specific nucleic acid sequences. These methods rely on the use of our proprietary SuperTracers to increase the specific activity of the probes that are then measured with supersensitive MPD instrumentation. SuperTracers coupled to DNA probes have been developed, and multiple coincidence methods to reduce biological background are being implemented. Preliminary experiments documented that the multiple coincidence probe method permits a considerable rejection of non- specific biological background. Overall, our studies confirmed that MPD permits improved qPCR methods and innovative direct methods of nucleic acid quantitation. Further development of these innovative methods are main topics for the proposed research, especially the development of the MPD enhanced direct quantitation of triplet repeats. PROPOSED COMMERCIAL APPLICATIONS: This project will develop procedures and supporting reagents which enable reliable direct detection and quantitation of low levels of DNA. We are developing robust direct detection methods which may be performed with minimal sample preparation in the presence of high backgrounds of indigenous DNA. Such an assay system would have wide application in a number of important research, development and 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. The commercial success drivers for these products are their unique performance for the quantitative detection of DNA/RNA at levels which cannot be achieved with competing technologies.