Project Brief
Tools For DNA Diagnostics (October 1994)Development of a Generic Technology for the Targeted Detection and Cleavage of DNA and RNADevelop simple-to-use, low-cost, diagnostic tools that rapidly detect specific DNA and RNA sequences for broad-based medical diagnosis and for tracking treatments. Sponsor: Third Wave Technologies, Inc.2800 S. Fish Hatchery RoadMadison, WI 53711
Third Wave Technologies, Inc., proposes to develop a tool box of faster, cheaper, and more user-friendly technologies for detecting and manipulating DNA and other nucleic acids as the basis for broad-based molecular diagnostic systems designed primarily for medical professionals. Specific applications of the technology include assessing tissues for transplantation suitability, forensic and paternity tests, diagnosing hereditary and infectious diseases, assessing susceptibility to specific diseases, and monitoring the response of disease pathogens to specific medical treatments. The basis for the proposed tool box derives from a set of deliberately modified bacterial enzymes called polymerases that fall under the company's trade name CleavaseTm, but that have yet to be developed into clinically usable diagnostic tools. The key principle behind the Cleavase enzymes is that they can easily be molecularly tuned to cleave any predetermined sequence of nucleic acids whether they be from healthy cells or pathogenic bacteria or viruses. A "reporter segment" built into each of the Cleavase-based systems sends out a signal when such a cleavage occurs. If the population of target DNA segments is high enough in a sample, for example, a thousand pathogenic viruses in a blood sample, the collective signal is strong enough for direct detection using "light" sensors. When there are too few copies of the target DNA in a sample for direct detection, the Cleavase Detection Reaction can circumvent the deficit. In this case, the first set of reporter segments triggers a series of molecular events that amplifies the amount of signal-emitting reporters up to detectable levels, even when there are as few as 10 target sequences in the sample. The technological challenge now is to develop these Cleavase technologies into extremely versatile, turn-key systems for clinical diagnosticians.
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