An Innovative, High-Throughput Method of SNP Haplotyping

该方案的目的是开发一种创新的、高通量、可靠和准确的SNP-单倍型方法。到目前为止，还没有高通量、可靠的SNP单倍型方法可用。这项建议的具体目的是开发一种新的、等位基因特异的寡核苷酸杂交方法，用于使用微阵列或384孔微滴定板格式确定单倍型。这些格式将允许高通量地确定包含两个SNP(微滴定板格式)或多个SNP(微阵列格式)的单倍型。加/减，杂交模式的自动评分将确定两条染色体的单倍型。以高通量方式准确地对SNP基因座进行单倍型的能力在以下方面有重要的应用：1)使用单倍型进行饱和基因分型研究，以缩小由先前与单个SNPs或其他标记进行的连锁研究所定义的候选基因组区域；2)连锁不平衡和关联研究位于候选基因组区域或通过其他方法确定的候选基因；3)分析大量样本以寻找先前确定的与特定疾病的易感性相关的单倍型；4)诊断测试和试剂盒，用于与疾病易感性、药物遗传学和免疫学简档相关的单倍型。拟议的商业应用：高通量、准确和可靠的SNP单倍型技术将为连锁不平衡和关联研究的群体分析提供支持。这类研究将允许对糖尿病、冠状动脉疾病和哮喘等复杂特征进行基因解剖。这对于了解和诊断许多常见疾病、开发诊断/预后检测试剂盒和发现新的治疗方法具有重要的商业应用价值。

The objective of this proposal is to develop an innovative, high- throughput, reliable and accurate method of SNP-haplotyping. To date, there are no high-throughput, reliable SNP-haplotyping methods available. The specific aims of this proposal are designed to develop a novel, allele-specific oligonucleotide hybridization approach for haplotype determination using either a microarray or a 384 well microtiter-plate format. These formats will allow the high-throughput determination of haplotypes containing 2 SNPs (microtiter-plate format) or multiple SNPs (microarray format) for both chromosomes of an individual. Plus/minus, automatable scoring of hybridization patterns will determine the haplotypes for both chromosomes. The ability to accurately haplotype SNP loci in a high-throughput fashion has important applications for; 1) saturation genotyping studies using haplotypes to narrow candidate genomic regions defined by previous linkage studies carried out with individual SNPs or other markers, 2) linkage disequilibriurn and association studies of genes located in candidate genomic regions or candidate genes identified by other methods, 3) analyzing large numbers of samples for haplotypes previously determined to be associated with the susceptibility of a particular disease, 4) diagnostic tests and kits for haplotypes associated with disease susceptibility, pharmacogenetic and immunologic profiling. PROPOSED COMMERCIAL APPLICATION: A high-throughput, accurate and reliable SNP-haplotyping technology will support population analyses for linkage disequilibrium and association studies. Such studies will allow the genetic dissection of complex traits such as diabetes, coronary artery disease and asthma. This has significant commercial application for the understanding and diagnosis of many common diseases, the development of diagnostic/prognostic test kits and the discovery of novel therapeutic treatments.