SBIR Phase I: Resonant Acoustic Cell Enrichment (RACE) Technique for Rare Cell Detection in Blood

这个小企业创新研究（SBIR）第一阶段项目解决了快速，成本效益和有效的样品制备分离稀有细胞（例如，病原体、循环肿瘤细胞等）大量的血液。将开发一种创新样品制备技术的原型，并在第一阶段展示其性能。共振声细胞富集（RACE？）该技术利用声电泳提供了一种连续流动、可自动化和快速的方法来替代目前用于分离稀有细胞的方法。RACE能够以10 s ml/min的速度处理大量血液，而不会对细胞活力产生不利影响。它允许基于将靶细胞结合到抗体包被的珠上并将珠快速浓缩到小体积的清洁缓冲液中来从全血中纯化稀有靶。使用这种方法，细胞可以在当前技术所需的一小部分时间内在优选的缓冲液中浓缩成小体积。因此，当用户等待时，靶细胞可以在几分钟内被分离。在第I阶段，将采用qPCR检查加标动物血样中细菌标志物的回收率和富集情况。拟议技术更广泛的商业影响是临床应用的潜力。例如，脓毒症是一种严重的医疗状况，通常在重症监护室治疗。治疗需要确定影响血流的感染类型。然而，病原体类型的鉴定受到每毫升血液中菌落形成单位数量较低（10 cfu/mL）的混淆。目前，这是补救的快速处理大量的血液（3-10毫升）和使用红细胞裂解，然后离心，以增加病原微生物的产量。使用RACE技术，病原体将被分离在小体积的干净缓冲液中，该缓冲液可以容易地使用遗传分析技术进行分析。拟议的样品制备系统的直接市场是基于DNA的即时基因检测。声泳装置将相对便宜;样品制备系统可以以5，000至10，000美元的价格推向市场。

This Small Business Innovation Research (SBIR) Phase I project addresses the need for rapid, cost effective and efficient sample preparation for isolating rare cells (e.g., pathogens, circulating tumor cells, etc.) from large blood volumes. A prototype of an innovative sample preparation technology will be developed and its performance will be demonstrated during Phase I. Resonant Acoustic Cell Enrichment (RACE?) technology employs acoustophoresis to provide a continuous flow, automatable, and rapid alternative to the current methods for isolation of rare cells. RACE enables processing of large blood volumes at 10s of ml/min without adversely affecting cell viability. It allows purification of rare targets out of whole blood based on binding target cells to antibody-coated beads and concentrating the beads rapidly into a small volume of clean buffer. Using this approach, cells can be concentrated into small volumes in a preferred buffer in a fraction of time required by current technology. Thus, target cells can be isolated in minutes while the user waits. During Phase I, recovery and enrichment of bacterial markers in spiked animal blood samples will be examined with qPCR. The broader commercial impact of the proposed technology is the potential for clinical applications. For example, sepsis is a serious medical condition that is usually treated in intensive care units. Treatment requires identification of the type of infection afflicting the blood stream. However, identification of the type of pathogen is confounded by low number of colony forming units per milliliters of blood (10 cfu/mL). Currently, this is remedied by the rapid processing of larger volumes of blood (3-10 mL) and the use of red blood cell lysis followed by centrifugation to increase the yield of pathogenic microorganisms. Using the RACE technology, the pathogens would be isolated in a small volume of clean buffer that can be readily analyzed using genetic analysis techniques. The immediate market for the proposed sample preparation system is point-of-care DNA-based genetic testing. Acoustophoresis devices will be relatively inexpensive; a sample preparation system could be delivered to the market for $5,000 to $10,000.