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从大量血液中分离稀有细胞（例如病原体、循环肿瘤细胞等）的快速、成本效益和高效样品制备的需求。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。共振声学细胞富集（RACE?）技术采用声电泳技术，为当前分离稀有细胞的方法提供连续流动、自动化和快速的替代方案。RACE能够以10s ml/min的速度处理大量血容量，而不会对细胞活力产生不利影响。它可以将靶细胞与抗体包被的小珠结合，并将小珠迅速浓缩到小体积的干净缓冲液中，从而从全血中纯化出罕见的靶标。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浓缩成首选缓冲液中的小体积。因此，当用户等待时，目标细胞可以在几分钟内分离出来。在第一阶段，将用qPCR检测加标动物血液样本中细菌标记物的恢复和富集。拟议技术的更广泛的商业影响是临床应用的潜力。例如，败血症是一种严重的疾病，通常在重症监护病房治疗。治疗需要确定影响血流的感染类型。然而，由于每毫升血液中菌落形成单位的数量较低（10 cfu/mL），使病原体类型的鉴定变得混乱。目前，通过快速处理大容量血液（3-10 mL）和使用红细胞裂解，然后离心以增加病原微生物的产量来补救这一问题。使用RACE技术，病原体将在小体积的干净缓冲液中分离出来，可以很容易地使用遗传分析技术进行分析。提议的样品制备系统的直接市场是基于dna的即时基因检测。声阻抗装置将相对便宜；样品制备系统可以以5000到10000美元的价格投放市场。

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.