Non-contact microchip PCR based on infrared Heating

描述（由申请人提供）：尽管目前流行的“基因组时代”即将结束（随着“后基因组时代”的到来），基因分析无疑将继续是一个多产的探索领域。遗传分析方法对于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（例如微卫星DNA）至关重要。因此，从原始生物样品中纯化核酸物质并通过聚合酶链反应（PCR）成功扩增目标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红外介导加热（即使用热灯）在微芯片中执行PCR的可行性，并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个样本）和单个微芯片上的DNA分离。PCR扩增将是完全非接触式的（红外加热和干涉测量温度传感），并能够在15分钟或更短的时间内完成DNA扩增（最多25个周期）。PCR产物的多通道电泳分析将随后在同一微芯片上进行，样品吞吐量超过每分钟一个样品（不到30分钟32个样品）。多重PCR与微设备分离的无缝集成将为诊断和其他各种领域提供具有“样品-答案-输出”能力的自动化仪器迈出一大步。这种方法不仅将利用微芯片技术的力量来减少样本量和提高PCR过程的速度，而且为真正集成的微芯片遗传分析奠定了基础（又名，真正的芯片实验室）。

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).