Development of Fluorescent Detector for DNA Sequencing

描述(由申请人提供)： 人类最常见的序列变异形式是单核苷酸多态(SNPs)，它可以影响蛋白质功能、基因的正确处理或影响基因的正常表达水平。我们建议开发一种新的荧光检测方法，并将其应用于信息丰富的SNPs的高通量识别，这可能导致对遗传病的更准确诊断，更好地预测风险易感性，或识别零星突变。所提出的技术被称为脉冲多线激励或PME。PME技术有两个潜在的优势，可以显著提高荧光灵敏度：(1)在基因组分析中对所有荧光团进行最佳激发；(2)“色盲”检测，它收集的光要多得多。这项技术与目前最先进的DNA测序仪器有很大的不同，目前DNA测序仪器具有单源激发和色散的特点，用于DNA序列鉴定。为了验证PME用于多色荧光检测的可行性，我们提出了构建单个毛细管面板，详细的检测实验来评估灵敏度，以及用于SNPs DNA测序的重构实验。 PME技术的成功实施将在临床诊断学、法医学和一般测序方法中的常规应用中得到广泛应用，并将具有为大多数人群进行定向序列变异分析的能力、灵活性和便携性。

DESCRIPTION (provided by applicant): The most commonly observed form of human sequence variation is single nucleotide polymorphisms (SNPs), which can affect protein function, proper processing of genes or affect the normal level of gene expression. We propose the development of a novel approach to fluorescence detection with application for high-throughput identification of informative SNPs, which could lead to more accurate diagnosis of inherited disease, better prognosis of risk susceptibilities, or identification of sporadic mutations. The proposed technology is called Pulse-Multiline Excitation or PME. The PME technology has two potential advantages, which could significantly increase fluorescence sensitivity: (1) optimal excitation of all fluorophores in the genomic assay and (2) "color-blind" detection, which collects considerably more light. This technology differs significantly from the current state-of-the-art DNA sequencing instrumentation, which features single source excitation and color dispersion for DNA sequence identification. To test the feasibility of PME for multi-color fluorescence detection, we propose the construction of a single capillary breadboard, detailed limit of detection experiments to assess sensitivity, and reconstruction experiments for DNA sequencing of SNPs. Successful implementation of the PME technology will have broad application for routine usage in clinical diagnostics, forensics, and general sequencing methodologies and will have the capability, flexibility, and portability of targeted sequence variation assays for a large majority of the population.