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

小型企业创新研究(SBIR)第一阶段项目旨在满足快速、经济高效地制备样品以分离稀有细胞(例如病原体、循环肿瘤细胞等)的需求。从大量的血液中。将开发一种创新的样品制备技术的原型，并将在第一阶段展示其性能。该技术采用声导引法，为当前稀有细胞分离方法提供了一种连续、自动化和快速的替代方法。RACE能够以10毫升/分钟的速度处理大容量血液，而不会对细胞活力产生不利影响。它允许从全血中提纯稀有靶标，方法是将靶细胞与抗体包裹的珠子结合，并将珠子快速浓缩到小体积的清洁缓冲液中。使用这种方法，电池可以在当前技术所需时间的一小部分内集中到首选缓冲器中的小体积中。因此，当用户等待时，目标小区可以在几分钟内被隔离。在第一阶段，将用定量聚合酶链式反应检测被添加的动物血液样本中细菌标志物的恢复和浓缩。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，由于每毫升血液中的菌落形成单位数较低(10cfu/ml)，病原体的类型鉴定受到了混淆。目前，这是通过快速处理更大数量的血液(3-10毫升)和使用红细胞裂解然后离心来增加病原微生物产量来补救的。使用RACE技术，病原体将被分离在少量干净的缓冲液中，可以很容易地使用遗传分析技术进行分析。拟议的样本制备系统的直接市场是基于护理点DNA的基因测试。声导设备将相对便宜；一个样品制备系统可能会以5000至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.