TEST FOR OCCULT MICROMETASTASES IN PRIMARY BREAST CANCER

建议开发和验证一种新的测定系统，以取代 现有的用于检测肿瘤中隐匿性微转移的免疫细胞化学测定法 原发性乳腺癌患者的骨髓。拟定含量 将提供简单、低成本和定量的终点。 该检测是基于一种创新的扩增机制，细菌 链反应（BCR），这是潜在的能够降低 检测限为1千万个骨髓细胞中的一个微转移。 BCR利用大肠杆菌的一种自养型菌株的活细胞 线圈以放大来自报告分子的信号，例如，酶标 克隆抗体在BCR条件下，报告分子 触发附近的自体互养细胞启动增殖链 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测定并评估其 使用模型系统的检测限。对于II期，BCR检测将 通过将其应用于探测和枚举神秘的 原发性乳腺癌患者骨髓标本中的微转移 癌最终目标是生产一种BCR检测试剂盒， 作为临床诊断工具。

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.