Chromogenic Her-2/neu Gene Amplification Assay

描述(申请人提供)：HER-2/neu蛋白过表达 和HER-2/neu基因的扩增是最被广泛接受的 侵袭性乳腺恶性行为的预后指标 癌症。HER-2/neu过表达的准确检测在 用人源化单抗鉴定适合进行血清疗法的患者 抗体(赫赛汀)。荧光原位杂交(FISH)已被发现 比免疫组织化学染色更准确。一种原位显色 建立了检测HER-2/neu基因扩增的杂交方法 石蜡包埋浸润性乳腺癌切片中的基因。与鱼不同， 这需要荧光光学，这在许多病理学中并不常见 实验室里，化验是在传统的Brightfield显微镜中读取的。 检测是基于自动金字学增强的纳米金，一种高度 提供密集黑色信号的灵敏方法，与通常的 用过的污渍。与FISH、组织学分级和组织学分级有良好的一致性 一系列101例乳腺癌的rnRNA杂交结果。我们现在提议 为17号染色体多切体、rnRNA表达或 伴癌蛋白检测作为解决边界问题的内部对照 放大案例。经过设计优化，它将被移植到一个 全自动玻片染色仪。一旦性能得到优化， 将进行多中心临床试验，以支持申请 上市前审批。

DESCRIPTION (provided by applicant): Overexpression of the Her-2/neu protein and amplification of the Her-2/neu gene are are the most widely accepted prognostic indicator of aggressive malignant behavior of invasive breast carcinoma. Accurate detection of Her-2/neu overexpression is critical in identifying patients suitable for serotherapy with humanized monoclonal antibody (Herceptin). Fluorescence in situ hybridization (FISH) has been found to be more accurate than immunohistochemical staining. A chromogenic in situ hybridization assay has been developed to detect amplification of the Her-2/neu gene in paraffin-embedded invasive breast carcinoma sections. Unlike FISH, which requires fluorescent optics not routinely available in many pathology laboratories, the assay is read in the conventional brightfield microscope. Detection is based on autometallographically enhanced Nanogold, a highly sensitive method giving a dense black signal, fully compatible with commonly used stains. Excellent concordance was found with FISH, histologic grading and mRNA:rnRNA hybridization for a series of 101 breast carcinomas. We now propose to add a secondary assay for chromosome 17 polysotomy, rnRNA expression or concomitant oncoprotein detection as an internal control to resolve borderline amplification cases. After design optimization, it will be ported to an automated slide staining instrument. Once performance is optimized, multi-center clinical trials will be conducted to support application for pre-market approval.