Diagnosing DNA Mutations Using Mismatch Repair Enzymes

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DESCRIPTION (provided by applicant): Trevigen/TreviMed has developed the Mutation Identification DNA Analysis System (MIDAS TM) that relies on DNA mismatch repair enzymes (MREs) for the detection and identification of known point mutations and pathogen DNA without the need for PCR amplification. The purpose of the research outlined in this Phase II proposal is to make MIDAS a robust and commercially viable clinical diagnostic product. To this end we propose the following specific aims: 1) increase the turnover rates of the MREs; 2) design MIDAS probes that incorporate donor/acceptor fluorophores which interact through fluorescence resonance energy transfer (FRET); and 3) optimize and clinically validate FRET-MIDAS reagents for the detection of high-risk oncogenic human papilloma virus (HPV) and the Coagulation factor V Leiden mutation, which are associated with cervical cancer and thrombophilia, respectively. Phase II support will lead to the development of a multiplexed, high-throughput, robust platform for detection of these proposed targets. We project that FRET-MIDAS will be able to accomplish all of this at a cost of $40 to $55 per test.