DETECTION OF K RAS MUTATIONS BY LIGASE CHAIN REACTION

通过连接酶链反应检测 K RAS 突变

• 批准号: 2009959
• 负责人: TERESA A LEHMAN
• 金额: $ 9.91万
• 依托单位: BIOSERVE BIOTECHNOLOGIES, LTD
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2009959
• 关键词: bioassay; biomarker; gene mutation; ligase; method development; oncogenes; polymerase chain reaction

The carcinogenic process is the result of accumulation of molecular damage to DNA, including activation of protooncogenes and mutation or deletion of tumor suppressor genes. Detection of the early alterations which may be present in various types of cancer could serve as sensitive biomarkers to facilitate early diagnosis of the disease. One of the most consistent and frequent genetic lesions in carcinogenesis is the activation mutation of the first base of codon 12 of the K-ras onconogene. An assay capable of detecting low levels of K-ras mutations could be used in the clinic for earlier diagnosis of premalignant changes. the ligase chain reaction (LCR) is a sensitive methodology which has only recently been utilized to detect human K-ras mutations. The specific aims of this project are to: (1) adapt assay to non-isotopic methods, (2) increase the number of reactions possible per analysis by using microtiter plates for the LCR, and (3) develop mulitiplexing analysis to simultaneously determine other sites of K-ras mutations. Technical innovations would be introduced to develop a screening assay or kit which would be introduced to develop a screening assay or kit which would be affordable, easy to perform and interpret, and rapid. Such an assay could provide significant health and economic benefits and have widespread commercial application. PROPOSED COMMERCIAL APPLICATIONS: The potential commercial application of Phase I and II of this project is the development of a non-isotopic assay using PCR-based LCR methodology to determine the K-ras status of human DNA samples. The procedure must be (1) accurate, (2) easy to perform and interpret, (3) sensitive, (4) rapid and (5) amenable to large scale screening.

致癌过程是分子积累的结果 DNA 损伤，包括原癌基因的激活和突变或 肿瘤抑制基因的缺失。 检测早期改变 可能存在于各种类型的癌症中，可以作为敏感的 生物标志物有助于疾病的早期诊断。 最有之一 致癌过程中持续且频繁的基因损伤是 K-ras 密码子 12 的第一个碱基的激活突变 癌基因。 能够检测低水平 K-ras 突变的检测方法 可用于临床早期诊断癌前病变 变化。 连接酶链式反应 (LCR) 是一种灵敏的方法 最近才被用来检测人类 K-ras 突变。 该项目的具体目标是：（1）使测定适应非同位素 方法，(2) 增加每次分析可能发生的反应数量 使用微量滴定板进行 LCR，以及 (3) 开发多重分析 分析以同时确定 K-ras 突变的其他位点。 将引入技术创新来开发筛选方法 或将被引入以开发筛选测定或试剂盒的试剂盒 这将是负担得起的、易于执行和解释并且快速的。 这种测定可以提供显着的健康和经济效益， 具有广泛的商业应用。 拟议的商业应用： 该项目一、二期潜在商业应用 是使用基于 PCR 的 LCR 的非同位素测定的开发 确定人类 DNA 样本 K-ras 状态的方法。 这 程序必须 (1) 准确，(2) 易于执行和解释，(3) 灵敏，(4) 快速，(5) 适合大规模筛查。