Chromogenic Her-2/neu Gene Amplification Assay

描述（由申请方提供）：Her-2/neu蛋白的过表达 Her-2/neu基因的扩增是最广泛接受的 浸润性乳腺癌侵袭性恶性行为的预后指标 carcinoma. Her-2/neu过表达的准确检测在 鉴定适合用人源化单克隆抗体进行血清治疗的患者 抗体（赫赛汀）。荧光原位杂交（FISH）已被发现 比免疫组化染色更准确。A原位显色 已经开发了杂交分析来检测Her-2/neu的扩增， 石蜡包埋的浸润性乳腺癌切片中的基因。与FISH不同， 其需要在许多病理学中常规不可用的荧光光学器件 在实验室中，在常规明场显微镜中读取测定。 检测是基于自动扫描增强纳米金，一个高度 灵敏的方法，提供密集的黑色信号，与通常的 用过的污渍与FISH、组织学分级和 101例乳腺癌的mRNA：mRNA杂交我们现建议 增加第二次检测17号染色体多聚体分裂、mRNA表达或 伴随癌蛋白检测作为内部对照，以解决边界 放大案例经过设计优化后，将移植到 自动载玻片染色仪。一旦性能得到优化， 将开展多中心临床试验，以支持申请 上市前批准。

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.