Self-Digitization Microfluidic Device for Digital PCR

用于数字 PCR 的自数字化微流控装置

• 批准号: 8645375
• 负责人: Jason Eugene Kreutz
• 金额: $ 15万
• 依托单位: LAMPROGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-18 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8645375
• 关键词: Address; Affect; Area; Biological; Biomedical Research; Cancer Diagnostics; Categories; Clinical; DNA; Development; Device Designs; Devices; Diagnostic; Disease; Drops; Future; Gene Expression; Gold; Image; Individual; Lead; Life; Measures; Medical; Methods; Microfluidic Microchips; Microfluidics; Modeling; Modification; Monitor; Nature; Nucleic Acids; Oils; Performance; Phase; Positioning Attribute; Production; Pump; RNA; Radial; Reaction; Research; Resolution; Sampling; Signal Transduction; Small Business Innovation Research Grant; Syringes; System; Technology; Validation; Variant; Viral Load result; Work; aqueous; clinical application; cost; cost effective; design; digital; flexibility; improved; instrument; instrumentation; meetings; optical disc; pathogen; pressure; prototype; public health relevance; scale up; statistics

DESCRIPTION (provided by applicant): This SBIR Phase I project will determine the feasibility of using Self-Digitization (SD) microfluidic technology as a commercially competitive nucleic acid quantification (NAQ) platform. Quantification of nucleic acids (both DNA and RNA) is utilized in various fields including general research, biomedical research, and clinical diagnostics, with specific applications in areas such as measuring gene expression levels, disease diagnostics including pathogen/viral load monitoring, cancer diagnostics, and more. Self-Digitization is a simple, robust technology that efficiently uses a network of channels and wells to spontaneously partition samples into an array of predefined well volumes. This platform is ideally suited for digital PCR (dPCR) applications. Digital PCR works by partitioning samples into thousands or even millions of individual volumes where each volume may or may not contain target DNA. Only volumes with target DNA give a positive signal resulting in a "digital" yes/no signal, from which Poisson statistics can then give direct determination of sample concentrations. Digital PCR directly provides absolute quantification, is robust against variations in reaction efficiency, is incredibly sensitive and has nearly unlimited resolution capabilities, making it technically superior to the current NAQ "gold standard" of qPCR. However, no commercial dPCR system is currently able to be economically competitive with qPCR. The objective of this proposal is to show the technical feasibility of a new centrifugal SD filling method within an optical disc (OD) style platform. The simplicity and flexibility of the SD system, its implementation within a method that enables parallelization and high-throughput experimentation, and the adaptation of existing commercialized components to achieve a complete platform will result in a cost-competitive product that will surpass existing commercial dPCR platforms, while also directly competing with qPCR systems. The approach will first validate the technical aspects of the platform by developing a thorough understanding of the loading/digitization mechanism, optimizing design features to maximize performance within a closed system, and to develop complete designs to meet needs in specific application categories. This will initially be carried out in scaled down versions to maximize iteration efficiency and then expanded to full prototype scale. Validation of this approach would then lead to future production of a mass producible device product and complete instrumentation setup. The product would be immediately useful for general biomedical research applications, and the workflow and nature of the closed system would also make it amenable to future clinical applications in a variety of medical fields.

描述（由申请人提供）：该SBIR I期项目将确定使用自数字化（SD）微流体技术作为商业竞争性核酸定量（NAQ）平台的可行性。核酸（DNA和RNA）的定量用于各种领域，包括一般研究、生物医学研究和临床诊断，具体应用于诸如测量基因表达水平、疾病诊断（包括病原体/病毒载量监测）、癌症诊断等领域。 自数字化是一种简单、强大的技术，它有效地使用通道和威尔斯网络，自发地将样品划分到预定义的孔体积阵列中。该平台非常适合数字PCR（dPCR）应用。数字PCR的工作原理是将样品分成数千甚至数百万个单独的体积，其中每个体积可能包含或不包含靶DNA。只有含有靶DNA的体积才能给出阳性信号，从而产生“数字”是/否信号，据此泊松统计可以直接测定样品浓度。数字PCR直接提供绝对定量，对变异具有鲁棒性 在反应效率方面，它非常灵敏，具有几乎无限的分辨率能力，使其在技术上上级目前的NAQ“黄金标准”qPCR。然而，目前没有商业化的dPCR系统能够与qPCR在经济上竞争。 本提案的目的是显示在光盘（OD）型平台内的新的离心SD填充方法的技术可行性。SD系统的简单性和灵活性， 其在能够并行化和高通量实验的方法中的实现，以及现有商业化组件的适应性以实现完整平台，将产生具有成本竞争力的产品，该产品将超过现有的商业dPCR平台，同时还直接与qPCR系统竞争。 该方法将首先通过对加载/数字化机制的透彻理解来验证平台的技术方面，优化设计功能以最大限度地提高封闭系统内的性能，并开发完整的设计以满足特定应用类别的需求。这将首先在缩小的版本中进行，以最大限度地提高迭代效率，然后扩展到完整的原型规模。这种方法的验证将导致未来大规模生产的设备产品和完整的仪器设置的生产。该产品将立即用于一般的生物医学研究应用，封闭系统的工作流程和性质也将使其适合未来在各种医学领域的临床应用。