DETECTION OF K RAS MUTATIONS BY LIGASE CHAIN REACTION

致癌过程是分子积累的结果， DNA损伤，包括原癌基因激活和突变，或 肿瘤抑制基因的缺失。 早期变化的检测 可能存在于各种类型的癌症中， 生物标志物，以促进疾病的早期诊断。 一个最 一致和频繁的遗传病变在癌变是 K-ras基因第12位密码子第一个碱基激活突变 致癌基因 一种能够检测低水平K-ras突变的检测方法 可用于临床早期诊断癌前病变 变化 连接酶链反应（LCR）是一种灵敏的方法 其最近才被用于检测人类K-ras突变。 本项目的具体目标是：（1）使分析适应非同位素 方法，（2）增加每次分析可能的反应数量， 使用LCR的微量滴定板，和（3）开发多重复合物 分析以同时确定K-ras突变的其他位点。 技术创新将被引入，以开发一种筛选分析方法， 用于开发筛选测定或试剂盒 这将是可负担的、易于执行和解释并且快速的。 这样的测定可以提供显著的健康和经济效益， 具有广泛的商业应用。 拟议的商业应用： 本项目一、二期的潜在商业应用 是开发一种非同位素检测方法， 方法来确定人类DNA样品的K-ras状态。 的 程序必须（1）准确，（2）易于执行和解释，（3） 灵敏，（4）快速和（5）适合大规模筛选。

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.