TEST FOR OCCULT MICROMETASTASES IN PRIMARY BREAST CANCER

建议开发和验证一个新的检测系统，以取代 现有的免疫细胞化学检测隐匿性微转移的方法 原发性乳腺癌患者的骨髓。建议的化验方法 将提供简单、低成本和量化的终点。 该检测是基于一种创新的放大机制，即细菌 链式反应(BCR)，它可能能够降低 检测极限为1000万个骨髓细胞中有一个微转移。 BCR利用了一种自养菌株的活细胞 线圈，用于放大来自报告分子的信号，例如，酶标记的 单克隆抗体。在BCR条件下，报告分子 触发附近的自体同步化细胞启动增殖链 反应导致离散的可见细菌菌落聚集在周围 肿瘤细胞。 第一阶段的目标是优化BCR分析并评估其 使用模型系统的检测限值。对于第二阶段，bcr检测将 通过将其应用于隐蔽的检测和枚举进行验证 原发性乳腺癌患者骨髓标本中的微转移 癌症。最终的目标是生产一种要投放市场的bcr检测试剂盒。 作为临床上的诊断工具。

It is proposed to develop and validate a new assay system to replace existing immunocytochemical assays for detecting occult micrometastases in bone marrow of patients with primary breast cancer. The proposed assay would offer simplicity, low cost, and a quantitative end point. The assay is based on an innovative amplification mechanism, the bacterial chain reaction (BCR), which is potentially capable of lowering the detection limit to one micrometastasis in 10 million bone marrow cells. The BCR utilizes living cells of an autosyntrophic strain of Escherichia coil to amplify signals from reporter molecules, e.g., enzyme-labeled monoclonal antibodies. Under the BCR conditions, reporter molecules trigger nearby autosyntrophic cells to initiate a proliferative chain reaction resulting in discrete visible bacterial colonies clustered around tumor cells. The objectives in Phase I are to optimize the BCR assay and assess its detection limits of using a model system. For Phase II, the BCR assay will be validated by applying it to detection and enumeration of occult micrometastases in bone marrow specimens from patients with primary breast cancer. The final objective is to produce a BCR assay kit to be marketed as a diagnostic tool in the clinic.