Correlative Chromogenic Gene and Protein assessment

相关显色基因和蛋白质评估

• 批准号: 6834406
• 负责人: RICHARD DENIS POWELL
• 金额: $ 17.53万
• 依托单位: NANOPROBES, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6834406
• 关键词: aneuploidy; bioimaging /biomedical imaging; breast neoplasm /cancer diagnosis; breast neoplasms; chromosomes; clinical research; diagnosis design /evaluation; epidermal growth factor; female; gene expression; gold; growth factor receptors; human tissue; image enhancement; immunocytochemistry; in situ hybridization; light microscopy; molecular probes; natural gene amplification; neoplasm /cancer genetics; protooncogene; women' s health

DESCRIPTION (provided by applicant): We propose a comprehensive HER2 status assay for gene amplification detection with concomitant protein assessment and an internal control for chromosome 17 polysomy in a single brightfleld light microscope slide staining protocol. Gene copy, protein overexpression, and the internal control will be marked by different colored stains. This will also enable visualization and counterstalning of the underlying tissue morphology, and provide a permanent record with no signal loss over time. In sltu hybridization detection by enzyme metallography has been combined with Immunohistochemical protein staining using Fast red K to give high-resolution, deady distinguished concomitant gene and protein staining. A centromeric ohromosome 17 probe will be added as an internal control for chromosome 17 polysomy, detected using either a second metallographic signal converted to a colored product using photographic dye coupling chemistry, or enzymatic deposition of a different colored substrate. After optimization of the staining and metaliographic reagents and protocols, the new assay will be evaluated in a parallel controlled perspective study: a sedes of 104 tumors, previously evaluated by lmmunohistochemistry and in situ hybridization using FISH, GOLDFISH and enzyme metallography, will be evaluated using the new assay. HER2 will be a model for the development of a general method applicable to many cancers and to the future development of other multi-analyte assays and staining procedures.

描述（由申请方提供）：我们提出了一种用于基因扩增检测的综合HER2状态试验，伴随蛋白质评估，并在单个Brightfield光学显微镜载玻片染色方案中使用17号染色体多体性的内部对照。基因拷贝、蛋白质过表达和内部对照将用不同颜色的染色剂标记。这也将实现底层组织形态的可视化和计数，并提供永久记录，随着时间的推移没有信号损失。在原位杂交检测中，用酶金相法结合免疫组织化学蛋白质染色法，用固红K进行染色，以获得高分辨率、精确区分的伴随基因和蛋白质染色。将添加着丝粒染色体17探针作为17号染色体多体性的内部对照，使用第二金相信号（使用照相染料偶联化学转化为有色产物）或不同有色底物的酶促沉积进行检测。在优化染色和金属学试剂和方案后，将在平行对照前瞻性研究中评价新测定法：将使用新测定法评价先前通过免疫组织化学和原位杂交使用FISH、GOLDFISH和酶金属学评价的104个肿瘤的种子。HER2将成为开发适用于许多癌症的通用方法以及未来开发其他多分析物测定和染色程序的模型。