Non-contact microchip PCR based on infrared Heating

基于红外加热的非接触式微芯片PCR

• 批准号: 6679896
• 负责人: James P Landers
• 金额: $ 34.36万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-20 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6679896
• 关键词: bioengineering /biomedical engineering; biomedical automation; biomedical equipment; clinical research; cyclization; genotype; heat stimulus; human tissue; infrared radiation; interferometry; microprocessor /microchip; neoplastic growth; nucleic acid amplification techniques; polymerase chain reaction; single nucleotide polymorphism; telemetry

DESCRIPTION (provided by applicant): Despite the current fad that the 'genome era' is coming to an end (as the 'post-genome era' gets underway), genetic analysis will undoubtedly continue to be a prolific field of exploration. Genetic analytical methods will be critical in fleshing out new mutations correlative of neoplastic growth, understanding the significance of single nucleotide polymorphisms, and developing better genotyping methods for, e.g., microsatellite DNA. Accordingly, the purification of nucleic acid material from crude biological samples and the successful amplification of target DNA sequences by the polymerase chain reaction (PCR) will remain a requisite protocol that is of vital importance. We have demonstrated the feasibility of executing PCR in microchips using IR-mediated heating (i.e., using a heat lamp) and have shown that this can be directly interfaced with separation on the same microchip. We now propose to further these developments by defining the appropriate microchip design and corresponding instrumentation for multiplexed PCR (36 samples at a time) and DNA separation on a single microchip. The PCR amplification will be completely non-contact (IR for heating and interferometry for temperature sensing), and capable of completing DNA amplification (up to 25cycles) in 15 minutes or less. Multi-channel electrophoretic analysis of the PCR products will ensue on the same microchip with a sample throughput of more than a sample per minute (32 samples in less than 30 minutes). The seamless integration of multiplexed PCR with separation on a microdevice will provide a giant step towards automated instrumentation with 'sample-in-answer-out' capabilities for diagnostics and a variety of other fields. Such an approach will not only exploit the power of microchip technology for reducing sample volumes and increasing the speed of the PCR process, but set the stage for truly integrated genetic analysis on a microchip (a.k.a., a true lab-on-a-chip).

描述（由申请人提供）：尽管目前流行的是“基因组时代”即将结束（随着“后基因组时代”的开始），遗传分析无疑将继续是一个多产的探索领域。遗传分析方法在充实与肿瘤生长相关的新突变、理解单核苷酸多态性的意义以及开发更好的基因分型方法方面至关重要，例如，微卫星DNA因此，从粗制生物样品中纯化核酸材料和通过聚合酶链式反应（PCR）成功扩增靶DNA序列将仍然是至关重要的必要方案。我们已经证明了使用IR介导加热在微芯片中执行PCR的可行性（即，使用加热灯），并且已经表明这可以直接与同一微芯片上的分离相连接。我们现在建议通过定义适当的微芯片设计和相应的仪器来进一步发展这些发展，用于多重PCR（一次36个样品）和在单个微芯片上分离DNA。PCR扩增将是完全非接触式的（红外加热和干涉温度传感），能够在15分钟或更短的时间内完成DNA扩增（最多25个循环）。PCR产物的多通道电泳分析将在同一个微芯片上进行，样品通量超过每分钟一个样品（32个样品在不到30分钟）。多重PCR与微设备分离的无缝集成将为诊断和各种其他领域的自动化仪器提供一个巨大的进步，具有“样品输入-回答”功能。这种方法不仅可以利用微芯片技术的力量来减少样品体积和提高PCR过程的速度，还可以为在微芯片上进行真正的集成遗传分析奠定基础。真正的芯片实验室）。