Development of a portable PME DNA sequencer

描述(申请人提供)：基因组科学的主要挑战之一是识别和了解单核苷酸多态(SNPs)在常见疾病中的作用。预计将开发新的技术，为准确诊断遗传病、确定导致常见疾病的风险因素以及确定患者的代谢特征提供必要的工具。这种技术在许多方面带来了预防的希望，例如延缓疾病发生或发展的预防性治疗，以及以最安全和最有效的结果进行药物治疗(S)。然而，目前，DNA测序技术还不足以在数千人中快速识别SNPs，更不用说我们社会的很大一部分人了。理想情况下，DNA测序技术将实时直接从一个人的基因组DNA中询问整个基因组序列，并在现场进行分析。因此，为了快速和准确地识别序列变异，迫切需要实时、便携设备的新技术，第二部分的解释和理解SNPs的作用超出了本提案的范围，但人们认为，对特定SNPs的发现，特别是代表风险因素或疾病病因的基因，将与拟议的研究并行进行。在这里，我们建议构建一种新的便携式DNA测序设备，具有直接从基因组DNA来源检测SNPs的潜力。为了实现这一点，我们描述了利用我们的脉冲多线激励(PME)技术与微流控分离芯片相结合的实验。与传统的DNA测序技术相比，MicroPME策略具有几个优势，从而利用紧凑和轻量的激光光源显著增强了荧光检测。为了测试便携式技术的可行性，我们设计了一种紧凑的PME探测器，该探测器连接到一个多通道微流控分离芯片，在一个手提箱足迹的范围内。该仪器将通过多重格式的已知SNPs的重建实验进行验证。可行性的论证将导致拟议的第二阶段应用，以开发集成的便携式DNA测序仪。这些研究计划的成功结果将意味着吞吐量和性能的显著提高、方法的简化和成本的降低。预计MicroPME仪器将在临床诊断、法医分析、军事应用和一般测序目的的常规使用中有广泛的应用。

DESCRIPTION (provided by applicant): One of the major challenges in genome sciences is to identify and understand the role of single nucleotide polymorphisrns (SNPs) in common disease. It is envisioned that new technologies will be developed in providing the necessary tools for accurate diagnosis of inherited disease, the determination risk factors contributing to common disease, and the identification of a patient's metabolic profile. Such technology brings promise in prevention in numerous ways such as prophylactic treatment to delay the onset or progression of disease, and treatment with medication(s) with the safest and most efficacious outcome. Presently however, DNA sequencing technology is not adequate in rapidly identifying SNPs in thousands of individuals, let alone a large portion of our society. Ideally, DNA sequencing technology would interrogate the entire genome sequence directly from a person's genornic DNA in realtime, and analyses would be done in the field. Thus, new technologies in real-time, portable devices are greatly needed for rapid and accurate identification of sequence variation, The second part of interpretation and understanding the role of SNPs is beyond the scope of this proposal, but it is believed that discoveries of specific SNPs in particular genes that represent risk factors or are causative agents of disease will be made in parallel with the proposed research. Here, we propose the construction of a new portable DNA sequencing device with the potential for detection of SNPs directly from genomic DNA sources. To accomplish this, we describe experiments utilizing our PuIse-Multiline Excitation (PME) technology coupled with microfluidic separation chips. The microPME strategy has several advantages over conventional DNA sequencing technology resulting in significantly enhanced fluorescent detection utilizing compact and light-weight laser sources. To test the feasibility of the portable technology, we purpose the construction of a compact PME detector coupled to a multi-channel microfluidic separation chip within the confinement of a suitcase footprint. The instrument will be validated by reconstruction experiments of known SNPs in a multiplex format. Demonstration of feasibility will result in a proposed phase II application to develop an integrated portable DNA sequencer. The successful outcome of these research plans would represent significant increases in throughput and performance, simplified methodologies, and reduced costs. It is anticipated that microPME instruments will have broad applications for routine usage for clinical diagnosis, forensic analyses, military applications, and general sequencing purposes.