Automated methods for rapidly purifying and concentrating bacteria from environmental interferents are needed in next-generation applications for anything from water purification to biological weapons detection. Though previous work has been performed by other researchers in this area, there is still a need to create an automated system that can both purify and concentrate target pathogens in a timely manner with readily available and replaceable components that could be easily integrated with a detection mechanism. Thus, the objective of this work was to design, build, and demonstrate the effectiveness of an automated system, the Automated Dual-filter method for Applied Recovery, or aDARE. aDARE uses a custom LABVIEW program that guides the flow of bacterial samples through a pair of size-based separation membranes to capture and elute the target bacteria. Using aDARE, we eliminated 95% of the interfering beads of a 5 mL-sample volume containing 10⁷ CFU/mL of E. coli contaminated with 2 µm and 10 µm polystyrene beads at 10⁶ beads/mL concentration., The target bacteria were concentrated to more than twice the initial concentration in 900 µL of eluent, resulting in an enrichment ratio for the target bacteria of 42 ± 13 in 5.5 min. These results show the feasibility and effectiveness of using size-based filtration membranes to purify and concentrate a target bacterium, in this case E. coli, in an automated system.

从水净化到生物武器检测的下一代应用都需要快速纯化和浓缩细菌免受环境干扰的自动化方法。尽管该领域的其他研究人员已经完成了之前的工作，但仍然需要创建一个自动化系统，该系统可以及时纯化和浓缩目标病原体，并且具有易于使用和可替换的组件，可以轻松地与检测机制集成。因此，这项工作的目标是设计、构建和演示自动化系统（应用恢复的自动双过滤方法或 aDARE）的有效性。aDARE 使用定制的 LABVIEW 程序引导细菌样品流过一对基于尺寸的分离膜以捕获和洗脱目标细菌。使用 aDARE，我们消除了 5 mL 样品体积中 95% 的干扰珠子，其中含有 10 7 ^CFU /mL 的大肠杆菌，这些大肠杆菌被 2 µm 和 10 µm 聚苯乙烯珠子以 10 ⁶珠子/mL 的浓度污染。在 900 µL 洗脱液中浓缩至初始浓度的两倍以上，5.5 分钟内目标细菌的富集率为 42 ± 13。这些结果表明，在自动化系统中使用基于尺寸的过滤膜来纯化和浓缩目标细菌（在本例中为大肠杆菌）的可行性和有效性。