Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, USA.
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, USA.
Lab Chip. 2022 Feb 1;22(3):621-631. doi: 10.1039/d1lc00865j.
The emergence and spread of multidrug resistant bacterial strains and concomitant dwindling of effective antibiotics pose worldwide healthcare challenges. To address these challenges, advanced engineering tools are developed to personalize antibiotic treatments by speeding up the diagnostics that is critical to prevent antibiotic misuse and overuse and make full use of existing antibiotics. Meanwhile, it is necessary to investigate novel antibiotic strategies. Recently, repurposing mono antibiotics into combinatorial antibiotic therapies has shown great potential for treatment of bacterial infections. However, widespread adoption of drug combinations has been hindered by the complexity of screening techniques and the cost of reagent consumptions in practice. In this study, we developed a combinatorial nanodroplet platform for automated and high-throughput screening of antibiotic combinations while consuming orders of magnitude lower reagents than the standard microtiter-based screening method. In particular, the proposed platform is capable of creating nanoliter droplets with multiple reagents in an automatic manner, tuning concentrations of each component, performing biochemical assays with high flexibility (, temperature and duration), and achieving detection with high sensitivity. A biochemical assay, based on the reduction of resazurin by the metabolism of bacteria, has been characterized and employed to evaluate the combinatorial effects of the antibiotics of interest. In a pilot study, we successfully screened pairwise combinations between 4 antibiotics for a model strain.
多药耐药菌株的出现和传播以及相应的有效抗生素的减少给全球医疗保健带来了挑战。为了应对这些挑战，开发了先进的工程工具，通过加快诊断来个性化抗生素治疗，这对于防止抗生素滥用和过度使用以及充分利用现有抗生素至关重要。同时，有必要研究新的抗生素策略。最近，将单种抗生素重新用于组合抗生素疗法已显示出治疗细菌感染的巨大潜力。然而，药物组合的广泛采用受到筛选技术的复杂性和试剂消耗成本的限制。在这项研究中，我们开发了一种组合纳米液滴平台，用于自动化和高通量筛选抗生素组合，同时消耗的试剂数量比标准的微孔板筛选方法少几个数量级。特别是，该平台能够自动生成具有多种试剂的纳升级液滴，调整每个组件的浓度，进行具有高灵活性（温度和持续时间）的生化测定，并实现高灵敏度检测。基于细菌代谢还原 Resazurin 的生化测定已得到表征，并用于评估感兴趣抗生素的组合效果。在一项试点研究中，我们成功地筛选了模型菌株中 4 种抗生素之间的两两组合。
