MEASURING DNA PROBE HYBRIDIZATIONS WITHOUT RADIOACTIVITY

在无放射性的情况下测量 DNA 探针杂交

• 批准号: 3491416
• 负责人: FLOYD E TAUB
• 金额: $ 5万
• 依托单位: DIGENE DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1987-04-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3491416
• 关键词: DNA; RNA; Retroviridae; autoradiography; chemical conjugate; defective virus; diagnostic tests; enzymes; gel electrophoresis; image processing; molecular biology; neoplasm /cancer diagnosis; oncogenes; oncogenic virus; peroxidases

Nucleic acid hybridization (NAH), an extremely important molecular biology research tool, has the potential to be a major diagnostic procedure. However, the current technique requires the use of expensive, hazardous and time consuming methods based on radioactive materials. Furthermore, NAH tests typically result in either non-quantitative or only semi-quantitative information, a serious limitation when it is important to know if an abnormal quantity of a sequence (e.g. oncogenes) is present. A new non-isotopic labeling procedure which takes less than 1 hour, requires less toxic and less expensive reagents, appears more reliable, and has sensitivity which is higher than previous methods has been implemented in preliminary experiments. An improved method for quantitating radioactive and non-isotopic hybridizations based upon computerized image processing has also been developed. A Phase I grant will permit us to: (1) test this labeling technique on nucleic acids from transformed cells to determine the feasibility of using this technique in clinical medicine; and (2) quantify the extent of hybridizations using a prototype image processing system. In Phase II, we will use these systems for the analysis and quantitation of oncogenes, potentially oncogenic viruses, and chemotherapy resistance in a variety of cell lives and clinical specimens.

核酸杂交（NAH），这是一种极为重要的分子生物学 研究工具有可能成为主要的诊断程序。 但是，当前的技术需要使用昂贵，危险和 基于放射性材料的耗时方法。 此外，不 测试通常会导致非定量或仅半定量 信息，一个严重的限制，重要的是要知道是否 存在异常的序列（例如癌基因）。 一个新的非同位标签程序，需要不到1小时， 需要较小的毒性和较低的试剂，看起来更可靠，并且 具有高于以前方法的灵敏度 在初步实验中。 改进的定量方法 基于计算机图像的放射性和非同位素杂交 也已经开发了处理。 我授予的阶段将允许我们：（1）测试此标签技术 来自转化细胞的核酸以确定使用的可行性 这项临床医学技术； （2）量化 使用原型图像处理系统进行杂交。 在第二阶段，我们将使用这些系统进行分析和定量 癌基因，潜在的致癌病毒和A中的化学疗法抗性 各种细胞生命和临床标本。