SBIR Phase II: Low-cost Detection and Enrichment of Nucleic Acids by Interfacing with Commercially Available Cell Sorters

SBIR 第二阶段：通过与市售细胞分选仪连接来低成本检测和富集核酸

• 批准号: 1556119
• 负责人: Adam Sciambi
• 金额: $ 74.87万
• 依托单位: Mission Bio, Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1556119&HistoricalAwards=false
• 关键词: SBIR; Phase; II; Low; cost

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be to enable affordable detection and enrichment of rare DNA or RNA molecules from a large background population. Currently, such detection is performed on expensive, dedicated instruments that have low sensitivity and are not able to enrich. The strategy is to encapsulate the fluorescently-identified target molecules in thin oil shells so that they mimic cells, and then process them on sensitive and high-throughput cell-sorting instruments that are commonly available. By offloading the detection and enrichment to such machines, all that is required by researchers is a simple tool that encapsulates the molecules in thin oil shells. Applications made possible by high sensitivity and enrichment are numerous, from rare pathogen or cell detection to target capture of uncommon mutant sequences with the goal of downstream sequencing. Between the reduced cost and expansion of capabilities, this approach is likely to become a widely adopted method.This SBIR Phase II project proposes to develop a deployable instrument that allows high-throughput, low-cost quantitation of DNA/RNA molecules in a sample by interfacing with existing flow cytometry instruments. The initial objective is to quickly engineer all the functionality desired in that final instrument into a rough alpha version. Its features will include reliable droplet generation from iteratively optimized hard-plastic microfluidic chips, and automation via an advanced fluid handling system with precise feedback control. The second objective is to build the actual beta version for customer trials. The beta version will replace most of the off-the-shelf parts with streamlined, custom engineering and focus on reducing size and cost while still improving function through more iterative chip optimization. The final objective is to use the beta instrument to optimize the protocols and reagents for several applications using customer samples and benchmarking. At the conclusion of the project, the goal is to have instruments at customer sites generating data.

这一小型企业创新研究(SBIR)项目的更广泛影响/商业潜力将是能够负担得起从大量背景人群中检测和浓缩稀有DNA或RNA分子。目前，这种检测是在昂贵的专用仪器上进行的，这些仪器灵敏度低，无法进行浓缩。其策略是将荧光识别的目标分子封装在薄的油壳中，使它们模仿细胞，然后在通常可用的灵敏和高通量的细胞分选仪器上处理它们。通过将检测和浓缩工作转移给这样的机器，研究人员所需要的只是一个简单的工具，将分子封装在薄薄的油壳中。高灵敏度和高浓缩使应用成为可能，从罕见的病原体或细胞检测到以下游测序为目标的不常见突变序列的靶向捕获。在降低成本和扩展能力之间，这种方法很可能成为一种被广泛采用的方法。这个SBIR第二阶段项目建议开发一种可部署的仪器，通过与现有的流式细胞仪接口，允许高通量、低成本地定量样本中的DNA/RNA分子。最初的目标是将最终仪器中所需的所有功能快速设计成一个粗略的阿尔法版本。它的特点将包括通过反复优化的硬塑料微流控芯片生成可靠的液滴，以及通过具有精确反馈控制的先进流体处理系统实现自动化。第二个目标是构建实际的测试版，供客户试用。测试版将用简化的定制工程取代大多数现成的部件，并专注于缩小尺寸和成本，同时仍通过更多迭代芯片优化来改进功能。最终目标是使用Beta工具，使用客户样本和基准来优化几个应用程序的协议和试剂。在项目结束时，目标是在客户现场使用仪器生成数据。