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型（SNP）在普通疾病中的作用。可以预见，将开发新技术，以提供准确诊断遗传疾病的必要工具，导致常见疾病的确定风险因素以及鉴定患者的代谢特征。这种技术以多种方式预防预防，例如预防治疗，以延迟疾病的发作或进展，并用最安全，最有效的结果进行药物治疗。但是，目前，DNA测序技术在迅速识别成千上万个个体的SNP中不足，更不用说我们社会的很大一部分了。理想情况下，DNA测序技术将直接从一个人的Genornic DNA实时询问整个基因组序列，并且将在该领域进行分析。因此，对于快速，准确地识别序列变化，非常需要实时，便携式设备的新技术，解释的第二部分和理解SNP的作用超出了该提案的范围，但是人们认为特定的发现是特定的发现。代表危险因素或疾病病因的特定基因的SNP将与拟议的研究并行。在这里，我们提出了一种新的便携式DNA测序装置的构建，该设备可能直接从基因组DNA源中检测SNP。为此，我们描述了使用我们的Puise-Multiniline激发（PME）技术以及微流体分离芯片的实验。与传统的DNA测序技术相比，MicroPME策略具有多个优势，从而通过紧凑和轻巧的激光源可显着增强荧光检测。为了测试便携式技术的可行性，我们目的是建造紧凑型PME探测器，并在限制手提箱足迹的情况下与多通道微流体分离芯片结合。该仪器将通过以多重格式的已知SNP进行重建实验来验证。可行性的证明将导致提出的II期应用程序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代表吞吐量和绩效，简化方法和降低成本的大幅提高。预计MicroPME仪器将在常规使用临床诊断，法医分析，军事应用和一般测序用途的常规使用情况下进行广泛的应用。