High Throughput System for FISH Analysis

最小残留病（MRD）目前通过病理检查、细胞遗传学、荧光激活细胞分选和聚合酶链反应（PCR）来识别。病理检查是评估癌细胞形态的标准方法，但检测限仅为10/-1至10/-2。PCR的检出限约为10/-5，但在扩增过程中存在定量不足和结果虚假的问题。荧光原位杂交技术（FISH）克服了现有方法敏感性和特异性的问题。改进的FISH探针的发展，如用于检测染色体易位的双融合探针，以及允许每秒分析多达1000个细胞的自动滑动扫描仪器，使FISH成为检测MRD中罕见癌细胞和监测治疗效果的理想方法。利用自动滑动扫描仪器和新型双融合FISH探针，第一阶段的SBIR旨在开发一种执行间期FISH分析的自动化系统。I期研究将使用t(14:18)（q32;q21）易位作为模型系统，这是大约60%的b细胞非霍奇金淋巴瘤的特征。第一阶段的研究将比较所提出的自动化方法与传统人工分析的灵敏度、可靠性和通量。拟议的商业应用：通过提供一种自动、快速、定量的检测罕见癌细胞的系统，拟议的系统将促进癌症的早期诊断和微小残留疾病的检测，并允许监测癌症治疗的疗效。

Minimum Residual Disease (MRD) is currently identified using pathologic examination, cytogenetics, fluorescence activated cell sorting, and polymerase chain reaction (PCR). Pathologic examination, which assesses cancer cell morphology, is the standard method, but the detection limit is only 10/-1 to 10/-2. PCR has a detection limit of approximately 10/-5, but suffers, but suffers from lack of quantitation and spurious results during the amplification process. Fluorescence in situ hybridization (FISH) overcomes problems of sensitivity and specificity of current methods. Development of improved FISH probes, such as dual fusion probes for detecting chromosomes translocations, and automated slide scanning instrumentation, which permits analysis of up to 1,000 cells per second, make FISH an ideal method for detecting rare cancer cells in MRD and monitoring the efficacy of treatment. Using automated slide scanning instrumentation and novel dual fusion FISH probes, this Phase I SBIR aims to develop an automated system for performing interphase FISH analysis. Phase I research will use the t(14:18)(q32;q21) translocation, which characterizes approximately 60% of B-cell Non-Hodgkin's Lymphoma, as a model system. Phase I research will compare the sensitivity, reliability and throughput of the proposed automated method with conventional manual analysis. PROPOSED COMMERCIAL APPLICATIONS: By providing an automated, rapid, quantitative system for detecting rare cancer cells, the proposed system will facilitate early diagnosis of cancer and detection of Minimal Residual Disease, and permit monitoring of the efficacy of cancer treatments.