Array Imaging digital PCR Platform

摘要 Combinati希望推出一款集成的阵列成像数字聚合酶链式反应平台 从实验室到市场的微流控耗材，以加快dPCR技术的采用。数位 聚合酶链式反应因其在基因组研究和临床研究领域的能力而受到关注。 检测罕见事件(高灵敏度)，不易受到抑制(高特异性)，不需要内部 标准(高精密度)，并可使用成熟的聚合酶链式反应试剂。然而，目前的dPCR仪器 都是资本密集型的，每个样本的成本很高，而且工作流程复杂。成功完成后 在我们的第一阶段目标中，我们将继续以下产品开发工作，将我们的平台带到 具有以下差异化功能的市场 ·从qPCR平稳过渡： O相应的工作流程：用户将试剂混合物添加到消耗品、试剂 数字化、热循环和实时成像垂直集成于一体 乐器。 O成本效益： ▪仪器：成像、热控和气动控制的进展 技术允许我们以大约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实现低单位数据成本 积分(&lt；$1)。 O开放分析化学：试剂将被分配在塑料微室中 一种专有乳化油。用户的qPCR配方可以直接转换到我们的 DPCR扩增平台。 O自动化兼容：标准板框允许用户自动化dPCR 使用液体处理机进行处理。 ·更强大和一致的数据： O热循环期间的实时成像：通过从预热背景中减去 从后热循环图像中提取循环图像，以及记录放大 每个分区的动态，用户只能计算显示PCR值的分区 尽量减少误报电话。 O每个数据点的固定分区数：泊松统计数据取决于 分区总数。我们的消耗品将为每个数据保证20K分区 点，不像ddPCR，它在11k到19k之间变化。 O已知体积：量化是通过将分子的数量除以总数来完成的 音量。我们将用共聚焦激光干涉法测量阵列的体积，以 为用户提供精确的音量以进行量化。

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.