School of Biomedical Engineering, McMaster University, Hamilton, Ontario, Canada.
Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.
Nat Commun. 2024 Jul 11;15(1):5626. doi: 10.1038/s41467-024-49710-2.
As bacteriophages continue to gain regulatory approval for personalized human therapy against antibiotic-resistant infections, there is a need for transformative technologies for rapid target identification through multiple, large, decentralized therapeutic phages biobanks. Here, we design a high throughput phage screening platform comprised of a portable library of individual shelf-stable, ready-to-use phages, in all-inclusive solid tablets. Each tablet encapsulates one phage along with luciferin and luciferase enzyme stabilized in a sugar matrix comprised of pullulan and trehalose capable of directly detecting phage-mediated adenosine triphosphate (ATP) release through ATP bioluminescence reaction upon bacterial cell burst. The tablet composition also enhances desiccation tolerance of all components, which should allow easier and cheaper international transportation of phages and as a result, increased accessibility to therapeutic phages. We demonstrate high throughput screening by identifying target phages for select multidrug-resistant clinical isolates of Pseudomonas aeruginosa, Salmonella enterica, Escherichia coli, and Staphylococcus aureus with targets identified within 30-120 min.
随着噬菌体在针对抗生素耐药性感染的个体化人类治疗方面不断获得监管批准,我们需要通过多个大型、分散的治疗性噬菌体生物库来实现快速目标识别的变革性技术。在这里,我们设计了一种高通量噬菌体筛选平台,该平台由一个便携式的单个货架稳定、即用型噬菌体文库组成,全部装在全包式固体片剂中。每个片剂都封装有一种噬菌体,以及稳定在由普鲁兰和海藻糖组成的糖基质中的荧光素和荧光素酶,该基质能够在细菌细胞破裂时通过 ATP 生物发光反应直接检测到噬菌体介导的三磷酸腺苷 (ATP) 释放。片剂组成还增强了所有成分的干燥耐受性,这应该使噬菌体更容易和更便宜地在国际间运输,从而增加治疗性噬菌体的可及性。我们通过在 30-120 分钟内识别选择的多药耐药性铜绿假单胞菌、肠炎沙门氏菌、大肠杆菌和金黄色葡萄球菌临床分离株的目标噬菌体,证明了高通量筛选的可行性。