Correlative Chromogenic Gene and Protein assessment

描述（由申请人提供）：我们提出了一种全面的HER2状态检测方法，用于基因扩增检测，同时进行蛋白质评估和17号染色体多体的内部控制，采用单亮眼光学显微镜载玻片染色方案。基因复制、蛋白过表达和内部控制将用不同颜色的染色标记。这也将使底层组织形态的可视化和反同步成为可能，并提供一个永久的记录，不会随着时间的推移而丢失信号。在sltu杂交检测酶金相结合免疫组织化学蛋白染色快速红K给出高分辨率，死亡区分伴随基因和蛋白染色。将添加一个着丝性17号染色体探针作为17号染色体多体的内部控制，使用第二金相信号转换为彩色产物，使用照相染料偶联化学或酶沉积不同颜色的底物进行检测。在对染色和金相分析试剂和方案进行优化后，新的检测方法将在平行对照视角研究中进行评估：先前通过免疫组织化学和FISH，金鱼和酶金相原位杂交评估的104个肿瘤将使用新的检测方法进行评估。HER2将成为开发一种适用于许多癌症的通用方法的典范，也将成为未来开发其他多分析物检测和染色方法的典范。

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.