Diagnosing DNA Mutations Using Mismatch Repair Enzymes

描述(由申请人提供)：Trevigen/TreviMed开发了突变识别DNA分析系统(MIDAS TM)，该系统依赖DNA错配修复酶(MRE)来检测和鉴定已知的点突变和病原体DNA，而不需要PCR扩增。这项第二阶段提案中概述的研究的目的是使Midas成为一种强大的和商业上可行的临床诊断产品。为此，我们提出了以下具体目标：1)提高MRE的周转率；2)设计MIDAS探针，整合通过荧光共振能量转移(FRET)相互作用的供体/受体荧光团；以及3)优化和临床验证FRET-MIDAS试剂，用于检测分别与宫颈癌和血栓形成相关的高危致癌人乳头瘤病毒(HPV)和凝血因子V Leiden突变。第二阶段的支持将导致开发一个多路复用、高吞吐量、强大的平台，用于探测这些拟议的目标。我们预计，FRET-MIDAS将能够完成所有这些任务，每次测试的成本为40至55美元。

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.