TELOMERIC DNA PROBES FOR CLINIC DIAGNOSTICS

这项研究的主要目标是开发一套DNA 探针试剂，以测定整个人类的端粒完整性 基因组染色体特异性端粒探针，与 着丝粒探针将成为临床上一套强大的试剂 染色体易位、末端缺失和融合的检测 人类遗传疾病和癌症的事件。多种分子 将采用各种技术来实现以下具体目标： (1)开发一套染色体特异性端粒DNA探针 优化用于荧光原位杂交（FISH）。试剂盒 建议用于多色单染色体分析和全基因组分析 分析. (2)为每个端粒区域， 微卫星和cRY（i）重复多态性（嘌呤- 个体之间的嘧啶重复DNA序列）。基于PCR的试剂盒 将开发包含150-200个多态性标记的引物 其将检测单亲二体性和亚端粒缺失， 与不明原因的智力迟钝有关的隐性易位， 流产和自然流产。多态性标记将是 用于自动化分析。染色体特异性端粒DNA 探针也将为人类基因组研究人员提供完成 染色体图谱和新的DNA探针和基因的区域定位。 拟议的商业应用：染色体特异性端粒DNA探针 将成为人类遗传性疾病的强有力的诊断试剂， 以染色体重排为特征的癌症。试剂盒将进行优化 荧光原位杂交和多重PCR- 基于人端粒完整性的自动化测定。

The primary objective of the proposed research is to develop a set of DNA probe reagents to assay telomere integrity throughout the entire human genome. Chromosome-specific telomeric probes, in combination with centromere probes, will be a powerful set of reagents for the clinical detection of chromosome translocations, terminal deletions, and fusion events in human genetic disease and cancer. A wide variety of molecular techniques will be employed to accomplish the following specific aims: (1) to develop a complete set of chromosome-specific telomeric DNA probes optimized for fluorescence in situ hybridization (FISH). Kits are proposed for multicolored single chromosome analysis and total genome analysis. (2) To develop, for each telomeric region, multiple microsatellite and cRY(i) repeat polymorphisms (variations in purine- pyrimidine repeat DNA sequences between individuals). A PCR based kit will be developed containing primers for 150-200 polymorphic markers which will detect uniparental disomy and subtelomeric deletions and cryptic translocations associated with unexplained mental retardation, miscarriage, and spontaneous abortion. The polymorphic markers will be multiplexed for automated analysis. Chromosome-specific telomeric DNA probes will also provide tools to human genome researchers for completion of chromosome maps and for regional mapping of new DNA probes and genes. PROPOSED COMMERCIAL APPLICATION: Chromosome-specific telomeric DNA probes will be powerful diagnostic reagents for human genetic diseases and cancers typified by chromosomal rearrangements. Kits will be optimized for multicolor fluorescence in situ hybridization and a multiplexed, PCR- based, automated assay for human telomere integrity.