A nanoluciferase-encoded bacteriophage illuminates viral infection dynamics of Pseudomonas aeruginosa cells.

IF 5.1 Q1 ECOLOGY ISME communications Pub Date : 2024-08-22 eCollection Date: 2024-01-01 DOI:10.1093/ismeco/ycae105
Sophia Zborowsky, Quentin Balacheff, Ioanna Theodorou, Rokhaya Kane, Raphaëlle Delattre, Joshua S Weitz, Régis Tournebize, Laurent Debarbieux
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Abstract

Bacteriophages (phages) are increasingly considered for both treatment and early detection of bacterial pathogens given their specificity and rapid infection kinetics. Here, we exploit an engineered phage expressing nanoluciferase to detect signals associated with Pseudomonas aeruginosa lysis spanning single cells to populations. Using several P. aeruginosa strains we found that the latent period, burst size, fraction of infected cells, and efficiency of plating inferred from fluorescent light intensity signals were consistent with inferences from conventional population assays. Notably, imaging-based traits were obtained in minutes to hours in contrast to the use of overnight plaques, which opens the possibility to study infection dynamics in spatial and/or temporal contexts where plaque development is infeasible. These findings support the use of engineered phages to study infection kinetics of virus-cell interactions in complex environments and potentially accelerate the determination of viral host range in therapeutically relevant contexts.

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纳米荧光素酶编码噬菌体照亮铜绿假单胞菌细胞的病毒感染动态。
由于细菌噬菌体(噬菌体)具有特异性和快速感染动力学,因此越来越多的人将其用于细菌病原体的治疗和早期检测。在这里,我们利用一种表达纳米荧光素酶的工程噬菌体来检测与铜绿假单胞菌溶解相关的信号,范围涵盖单细胞到群体。利用几种铜绿假单胞菌菌株,我们发现从荧光强度信号推断出的潜伏期、爆发大小、受感染细胞的比例和培养效率与传统种群检测的推断结果一致。值得注意的是,与使用隔夜斑块相比,基于成像的特征可在数分钟至数小时内获得,这为在斑块发育不可行的空间和/或时间背景下研究感染动态提供了可能性。这些发现支持使用工程噬菌体来研究复杂环境中病毒-细胞相互作用的感染动力学,并有可能加快确定治疗相关情况下的病毒宿主范围。
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