High Throughput System for FISH Analysis

最小残留病(MRD)目前通过病理学检查、细胞遗传学、荧光激活的细胞分选和聚合酶链式反应(PCR)来确定。病理检查是评估癌细胞形态的标准方法，但检测下限仅为10/-1至10/-2。聚合酶链式反应的检测下限约为10/-5，但在扩增过程中存在定量不足和假结果的问题。荧光原位杂交(FISH)克服了目前方法的敏感性和特异性问题。改进的FISH探针的开发，如用于检测染色体易位的双融合探针，以及每秒可分析多达1,000个细胞的自动载玻片扫描仪器，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。使用自动载玻片扫描仪器和新型双融合FISH探针，第一阶段SBIR旨在开发一种执行相间FISH分析的自动化系统。第一阶段的研究将使用t(14：18)(q32；q21)易位作为模型系统，该易位表现了大约60%的B细胞性非霍奇金淋巴瘤。第一阶段的研究将对拟议的自动化方法的灵敏度、可靠性和吞吐量与传统的人工分析进行比较。拟议的商业应用：通过提供自动、快速、定量的检测罕见癌症细胞的系统，拟议的系统将有助于癌症的早期诊断和微小残留病的检测，并允许监测癌症治疗的效果。

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.