Development of a portable PME DNA sequencer

便携式PME DNA测序仪的开发

• 批准号: 6796095
• 负责人: Michael L. Metzker
• 金额: $ 30.71万
• 依托单位: LASERGEN
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-07 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6796095
• 关键词: DNA; bioengineering /biomedical engineering; biomedical equipment development; computer data analysis; computer program /software; computer system design /evaluation; fluid flow; fluorescence; genome; method development; nucleic acid sequence; portable biomedical equipment; single nucleotide polymorphism

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.

描述（由申请人提供）：基因组科学的主要挑战之一是识别和理解单核苷酸多态性（SNP）在常见疾病中的作用。据设想，将开发新技术，为准确诊断遗传性疾病、确定导致常见疾病的风险因素以及识别患者的代谢谱提供必要的工具。这种技术以多种方式为预防带来了希望，例如预防性治疗以延迟疾病的发作或进展，以及使用具有最安全和最有效结果的药物进行治疗。然而，目前，DNA测序技术还不足以快速识别数千人的SNP，更不用说我们社会的大部分人了。理想情况下，DNA测序技术将直接从一个人的基因组DNA中实时询问整个基因组序列，并在现场进行分析。因此，非常需要用于快速和准确识别序列变异的实时便携式设备的新技术。解释和理解SNP的作用的第二部分超出了本提案的范围，但据信，在代表风险因素或疾病病原体的特定基因中发现特定SNP将与所提议的研究并行进行。在这里，我们提出了一种新的便携式DNA测序设备的建设与直接从基因组DNA来源的SNPs检测的潜力。为了实现这一点，我们描述了利用我们的脉冲多线激发（PME）技术与微流控分离芯片的实验。与传统的DNA测序技术相比，microPME策略具有几个优点，从而利用紧凑且重量轻的激光源显著增强了荧光检测。为了测试的便携式技术的可行性，我们的目的是一个紧凑的PME检测器耦合到一个多通道的微流控分离芯片内的手提箱足迹的局限性的建设。该仪器将通过以多重格式对已知SNP进行重建实验来验证。可行性论证将导致拟议的第二阶段应用程序，以开发一个集成的便携式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着提高、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用途中具有广泛的应用。