DETECTING GENE DOSAGE MUTATIONS WITH A PCR-BASED METHOD

基因剂量异常可能发生在基因水平(如地中海贫血)、亚染色体水平(如连续基因综合征)或染色体水平(非整倍体)。本项目的目标是开发检测常见非整倍体和相邻基因综合征的分子诊断试剂盒。Genaco的研究人员已经开发出一种方法，可以高通量检测基因剂量变化。关键是利用精心设计的基于聚合酶链式反应的定量系统。将构建一个“参考基因组”，其中包含每个被测试座位的对照等位基因的副本。当与基因组DNA共扩增时，参考基因组将有助于一次测量多个基因座的相对基因剂量。该项目的最终产品可能是用于快速准确检测三体(21、18、13三体和其他)的诊断试剂盒和用于检测常见连续基因综合征的试剂盒(如DeGeorge、Prader-Willi/Angelman、Miller-Dieker、Smith-Magenis、Rubinstein-Taybi、Langer-Giedion、Beckwith-Wiedemann和WAGR等)。建议的商业应用：可以根据建议的研究开发分子诊断试剂盒。一种快速、准确的基因剂量分析方法可用于染色体非整倍体(包括唐氏综合征)、微缺失综合征(包括William综合征、DiGeorge综合征等)和癌症特异性等位基因丢失的检测。

Gene dosage abnormalities may occur at gene level (such as a thalassemias), subchromosome level (such as contiguous gene syndromes), or chromosome level (aneuploidy). The objective of this project is to develop molecular diagnostic kits for thedetection of common aneuploidies and contiguous gene syndromes. A methodology has been developed by Genaco's researchers that allows high throughput detection of gene dosage alterations. The key is to utilize a carefully designed PCR based quantitation system. A "reference genome" will be constructed that contains a copy of control allele for each tested locus. When co-amplified with genomic DNA, the reference genome will help to measure the relative gene dosage of many loci at a time. The end products of the project may be a diagnostic kit for fast and accurate detection of trisomies (trisomies 21, 18, 13 and others) and a kit for the detection of common contiguous gene syndromes (such as DeGeorge, Prader-Willi/Angelman, Miller-Dieker, Smith-Magenis, Rubinstein-Taybi, Langer-Giedion, Beckwith-Wiedemann and WAGR etc.) PROPOSED COMMERCIAL APPLICATIONS: Molecular diagnostic kits can be developed based on the proposed research. A fast and accurate gene dosage analysis method could be used for the detection of chromosome aneuploidy (including Down's syndrome), microdeletionsyndromes (including William syndrome, DiGeorge Syndrome, etc), and cancer specific allelic loss.