Nanodroplet PCR Lab-Chip for Clinical Pathogen Detection

描述(申请人提供)：快速、准确地检测和鉴定病原体对于有效管理和治疗传染病至关重要。虽然包括聚合酶链式反应在内的分子诊断技术是一种有希望的快速和灵敏的病原体检测方法，但它们存在一些技术缺陷，这些缺陷减缓了它们在临床上的接受程度。微流控芯片实验室技术通过大幅提高自动化水平、吞吐量和可靠性，同时降低分析成本和仪器尺寸，有望克服其中许多限制。我们的长期目标是开发一个高度灵活的芯片实验室平台，以我们创新的基于纳升液滴的“数字微流控”概念为基础，通过实时PCR检测病原体。最终，我们设想这个平台将使新一代高度自动化的手持仪器能够为传染病的诊断提供快速、准确和多元化的分子检测结果。为了实现这一目标，在第一阶段，我们将展示在我们的平台上进行并行自动化实时聚合酶链式反应的可行性。我们将开发一种原型芯片，能够以完全可编程和组合的方式自动建立纳米液滴反应。我们将进一步证明，薄膜加热器可以直接集成到芯片中来执行热循环操作。在以后的阶段，探测器和控制电子设备也将被微型化或集成到芯片上。此外，在第二阶段，我们计划将我们的原型设备应用于极低出生体重婴儿的念珠菌检测问题，并通过参与与杜克大学合作的多中心临床研究来严格验证我们的技术。

DESCRIPTION (provided by applicant): Rapid and accurate detection and identification of pathogens is crucial for the effective management and treatment of infectious disease. While molecular diagnostic techniques including PCR are a promising approach for rapid and sensitive detection of pathogens, they suffer from a number of technical drawbacks which have slowed their acceptance in the clinic. Microfluidic lab-on-a-chip technologies are well-poised to overcome many of these limitations by providing vastly increased levels of automation, throughput and reliability while reducing assay costs and instrument sizes. Our long term goal is to develop a highly-flexible lab-on-a-chip platform for detection of pathogens by real-time PCR based on our innovative nanoliter droplet-based "digital microfluidic" concept. Ultimately, we imagine that this platform would enable a new generation of highly-automated handheld instruments capable of providing fast, accurate, and multiplexed molecular test results for the diagnosis of infectious disease. Towards this goal, in phase I, we will demonstrate the feasibility of performing parallel automated real-time PCR reactions on our platform. We will develop a prototype chip capable of automatically setting-up nanodroplet reactions in a fully-programmable and combinatorial fashion. We will further demonstrate that thin-film heaters can be integrated directly into the chip to perform the thermocycling operations. In later phases the detector and control electronics would also be miniaturized or integrated onto the chip. Additionally, in phase II we propose to apply our prototype device to the problem of Candida detection in extremely low-birth-weight infants and rigorously validate our technology through participation in a multi-center clinical study in collaboration with Duke University.