MEASURING DNA PROBE HYBRIDIZATIONS WITHOUT RADIOACTIVITY

核酸杂交是一种极其重要的分子生物学 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、危险和 基于放射性材料的耗时方法。再者，不是的 测试通常导致非定量或仅半定量的结果 信息，这是一个严重的限制，当知道是否有 存在异常数量的序列(例如癌基因)。 一种新的非同位素标记程序，耗时不到1小时， 需要毒性更低、价格更低的试剂，看起来更可靠，而且 具有比以前的方法更高的灵敏度 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 加工技术也得到了发展。 第一阶段拨款将允许我们：(1)在 从转化细胞中提取核酸以确定使用 临床医学中的这项技术；以及(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来分析和量化 癌基因、潜在致癌病毒和化疗耐药性 细胞生命和临床标本的多样性。

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.