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.

核酸杂交是一种极其重要的分子生物学技术 研究工具，有可能成为一个主要的诊断程序。 然而，目前的技术需要使用昂贵的、危险的和有害的化学物质。 基于放射性物质的耗时方法。 此外，NAH 测试通常导致非定量或仅半定量 信息，一个严重的限制，当重要的是要知道， 存在序列（例如癌基因）的异常量。 一种新的非同位素标记程序，需要不到1小时， 需要更少的毒性和更便宜的试剂，看起来更可靠， 灵敏度高于已实施的先前方法 在初步实验中。 一种改进的定量方法 基于计算机图像的放射性和非同位素杂交 加工也得到了发展。 第一阶段的赠款将允许我们：（1）测试这种标记技术， 从转化的细胞中提取核酸，以确定使用 该技术在临床医学中的应用;（2）量化 杂交使用原型图像处理系统。 在第二阶段，我们将使用这些系统分析和定量 致癌基因、潜在致癌病毒和化疗耐药性， 各种细胞生命和临床标本。