Innovative molecular cytogenetic techniques for early leukemia/lymphoma detection

用于早期白血病/淋巴瘤检测的创新分子细胞遗传学技术

• 批准号: 6992624
• 负责人: JOE N LUCAS
• 金额: $ 23.18万
• 依托单位: CHROMOTRAX, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6992624
• 关键词: chromosome translocation; cytogenetics; diagnosis design /evaluation; early diagnosis; flow cytometry; fluorescent dye /probe; immunomagnetic separation; leukemia; lymphoma; neoplasm /cancer diagnosis; neoplasm /cancer genetics; nucleic acid hybridization; nucleic acid probes; nucleic acid repetitive sequence; subtraction hybridization; technology /technique development

DESCRIPTION (provided by applicant): We propose an innovative approach combining two separate, novel techniques to greatly enhance accurate detection of leukemia or lymphoma cells, which can be used ultimately for diagnostics and early detection of minimal residual disease (MRD) in bone marrow, lymph nodes, and peripheral blood. The novel techniques of our approach are: (1) A two-step method to generate repetitive sequences depleted DMA probes. The method includes subtracting hybridized biotin-labeled repetitive-sequence DNA complex with phenol and chloroform and a second subtraction using avidin labeled magnetic beads. The unique sequences are then recovered using our dual unique sequence primers. (2) A novel technique of hybridizing chromosomes in suspension with fluorescently-labeled repetitive sequences depleted DNA probes, in combination with flow cytometric analysis or magnetic sorting, in order to sensitively, precisely and rapidly quantify leukemia or lymphoma-related chromosome translocations and rearrangements. Morphologic examination has a detection sensitivity of 1 in 100. PCR technique is very sensitive, but lack of accurate quantitation still is a problem. By contrast, hybridizing chromosomes in suspension has a potential sensitivity of 1 in 1,000,000. We propose to use leukemia as a model to demonstrate solution hybridization technique in conjunction with repetitive sequences depleted DNA probes as a sensitive and specific technique for accurate detection of MRD. Early detection of MRD will increase patient survival and profoundly impact cancer patient management.

描述（由申请人提供）：我们提出了一种创新方法，结合了两种独立的新型技术，以极大地增强对白血病或淋巴瘤细胞的准确检测，最终可用于诊断和早期检测到骨髓，淋巴结和外围血液的最小残留疾病（MRD）。我们方法的新技术是：（1）一种两步方法，用于生成重复序列耗尽的DMA探针。该方法包括使用苯酚和氯仿减去杂交生物素标记的重复序列DNA复合物，以及使用Avidin标记的磁珠的第二次减法。然后使用我们的双唯一序列引物恢复唯一的序列。 （2）一种在悬浮液中杂交染色体的新技术，与荧光标记的重复序列耗尽的DNA探针结合了流式细胞仪分析或磁分析，以敏感，精确，快速地量化白血病或淋巴瘤相关的染色体易月化染色体转运和重新分析。 形态检查的检测灵敏度为100中的1个。PCR技术非常敏感，但缺乏准确的定量仍然是一个问题。相比之下，悬浮液中的杂交染色体的潜在灵敏度为1,000,000。我们建议将白血病作为模型，以与重复序列结合使用溶液杂交技术，耗尽DNA探针作为一种敏感和特定技术，以准确检测MRD。早期发现MRD将增加患者的生存率，并对癌症患者管理产生深远影响。