Diagnosing DNA Mutations Using Mismatch Repair Enzymes

使用错配修复酶诊断 DNA 突变

• 批准号: 6730644
• 负责人: LEONARD S BAZAR
• 金额: $ 41.88万
• 依托单位: TREVIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6730644
• 关键词: DNA repair; cell line; coagulation factor V; colorectal neoplasms; enzyme activity; fluorescence resonance energy transfer; genetic disorder diagnosis; human genetic material tag; human papillomavirus; human subject; molecular cloning; nucleic acid quantitation /detection; phosphoester ligase; point mutation; protein purification; technology /technique development; thermophilic organism

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.

描述（由申请人提供）：Trevigen/TreviMed开发了突变鉴定DNA分析系统（MIDAS TM），该系统依赖于DNA错配修复酶（MRE）检测和鉴定已知的点突变和病原体DNA，而无需PCR扩增。本II期提案中概述的研究目的是使MIDAS成为一种强大且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）增加MRE的周转率：2）设计掺入供体/受体荧光团的MIDAS探针，所述供体/受体荧光团通过荧光共振能量转移（FRET）相互作用; 3）优化并临床验证FRET-MIDAS试剂检测高危致癌人乳头瘤病毒（HPV）和凝血因子V Leiden突变，它们分别与宫颈癌和血栓形成倾向有关。第二阶段的支持将导致开发一个多路复用，高通量，强大的平台，用于检测这些拟议的目标。我们预计，FRET-MIDAS将能够以每次测试40至55美元的成本完成所有这些。