Array Imaging digital PCR Platform

阵列成像数字PCR平台

• 批准号: 9768589
• 负责人: Paul Ju-Sung Hung
• 金额: $ 60万
• 依托单位: COMBINATI, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-15 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9768589
• 关键词: Adoption; Air; Attention; Automation; Biological Assay; Biological Markers; Capital; Chemistry; Clinical Research; Collaborations; Color; Comb animal structure; Communities; Complement; Computer software; Consultations; Cost efficiency; Data; Development; Devices; Diagnosis; Dyes; Emulsions; Enzyme Activation; Event; Genetic; Genetic Markers; Genomics; Goals; Image; Image Analysis; Industrialization; Injections; Interferometry; Lasers; Legal patent; Letters; Liquid substance; Malignant Neoplasms; Market Research; Measures; Microfluidics; Molds; Monitor; Mutation; Mutation Detection; Nucleic Acids; Oils; Optics; Performance; Phase; Process; Production; Reagent; Recipe; Research; Running; Sampling; San Francisco; Slide; Small Business Innovation Research Grant; Specificity; Statistical Data Interpretation; Statistical Models; System; Technology; Temperature; Testing; Time; Treatment Efficacy; Viral Load result; cell free DNA; commercialization; cost; design; digital; digital imaging; disorder prevention; experimental study; fluorescence imaging; graphical user interface; image processing; imaging Segmentation; imaging capabilities; imaging platform; improved; innovation; instrument; light intensity; off-patent; operation; outcome forecast; pressure; product development; prototype; rare cancer; real-time images; sensor; statistics

Abstract Combinati is looking to bring an integrated Array Imaging digital PCR platform with injection molded microfluidic consumables from lab to marketplace to accelerate the adoption of dPCR technology. Digital PCR has drawn attentions in both genomic research and clinical research communities for its ability to detect rare events (high sensitivity), less prone to inhibition (high specificity), no need for an internal standard (high precision), and access to well-developed PCR reagents. However, current dPCR instruments are capital intensive, cost per sample is high, and the workflow is complicated. With successful completion of our Phase 1 aims, we will continue the following product development effort to bring our platform to market with the following differentiated features • Smooth transition from qPCR: o Equivalent workflow: The user will add the reagent mix to the consumable, reagent digitization, thermal cycling, and real-time imaging is vertically integrated into one instrument. o Cost efficiency: ▪ Instrument: The advance in imaging, thermal control and pneumatic control technologies allow us to sell the instrument at ~$30k, similar to qPCR instruments. ▪ Consumable: The scalable injection molding technology enables low cost per data point (<$1). o Open assay chemistry: The reagents will be partitioned in plastic microchambers instead of proprietary emulsion oil. The user’s qPCR recipe can be directed transitioned to our dPCR platform. o Automation compatible: The standard plate frame allows the user to automate the dPCR process with liquid handlers. • More robust and consistent data: o Real-time imaging during thermal cycling: By subtracting from background pre-thermal cycling image from the post-thermal cycling image, as well as recording amplification dynamics of each partition, the user is able to count only partitions showing PCR to minimize false positive calls. o Fixed number of partitions for every data point: Poisson statistics is dependent on the total number of partitions. Our consumable will guarantee 20k partitions for each data point, unlike ddPCR, which varies between 11k to 19k. o Known volume: Quantification is done by dividing the number of molecules to the total volume. The volume of our array will be measured with confocal laser interferometry to give the user precise volume for quantification.

摘要 Combinati希望推出一个集成的Array Imaging数字PCR平台， 从实验室到市场的微流控耗材，以加速dPCR技术的采用。数字 PCR由于其在基因组研究和临床研究领域的能力而引起关注。 检测罕见事件（高灵敏度），不易抑制（高特异性），无需内部检测 标准（高精度），并获得完善的PCR试剂。然而，目前的dPCR仪器 是资本密集型的，每个样品的成本高，并且工作流程复杂。成功完成 我们的第一阶段目标，我们将继续以下产品开发工作，使我们的平台， 具有以下差异化特征的市场 ·从qPCR平滑过渡： 等同的工作流程：用户将试剂混合物添加到耗材、试剂盒和试剂盒中。 数字化、热循环和实时成像垂直集成为一体 仪器 成本效率： 仪器：成像、热控和气动控制的进步 技术使我们能够以约3万美元的价格出售该仪器，类似于qPCR仪器。 耗材：可扩展的注塑技术可降低每数据成本 点（<$1）。 开放式检测化学：试剂将在塑料微室中分配， 专利乳化油。用户的qPCR配方可以直接转换到我们的 dPCR平台。 o自动化兼容：标准板框架允许用户自动化dPCR 用液体处理器处理。 ·更可靠和一致的数据： 热循环期间的实时成像：通过从背景中减去预热温度， 从热循环后的图像循环图像，以及记录放大 每个分区的动态，用户能够只计数显示PCR的分区， 最大限度地减少假阳性呼叫。 o每个数据点的固定分区数：泊松统计取决于 分区总数。我们的耗材将保证每个数据20k分区 与ddPCR不同，ddPCR在11k到19k之间变化。 o已知体积：通过将分子数量除以总数来进行量化 音量.我们的阵列的体积将用共焦激光干涉测量法测量， 为用户提供精确的定量体积。