Department of Biology, Institute of Microbiology, ETH Zurich, Zürich, Switzerland.
Methods Mol Biol. 2021;2357:253-272. doi: 10.1007/978-1-0716-1621-5_17.
A previously unappreciated link between persisters and the emergence and spread of antibiotic resistance has been recently established. The bulk of this research has been conducted in vitro, but some studies are beginning to elucidate the importance of persister reservoirs in both antibiotic treatment failure and the spread of antibiotic resistance using in vivo models. In order to further this research, careful analyses of the mechanisms of persister reservoir formation as well as the dynamics of persister survival and postantibiotic regrowth are of importance. Here, we present a mouse model to quantitatively study Salmonella persisters in vivo. By using neutral unique sequence barcodes, we describe the quantitative analysis of rare events (aka bottlenecks) associated with persister reservoir formation, survival, and reseeding of the gut lumen. This provides quantitative data for persister-fueled plasmid transfer in vivo. Although this chapter describes analysis of Salmonella persisters in a mouse model, these concepts can be applied to any experimental system provided that tractable experimental systems are present.
最近发现了持久性细菌与抗生素耐药性的产生和传播之间以前未被重视的联系。这项研究的大部分是在体外进行的,但一些研究开始阐明持久性细菌库在抗生素治疗失败和抗生素耐药性传播中的重要性,使用体内模型。为了进一步开展这项研究,仔细分析持久性细菌库形成的机制以及持久性细菌的生存和抗生素后再生动力学非常重要。在这里,我们提出了一种小鼠模型来定量研究体内沙门氏菌持久性细菌。通过使用中性独特序列条形码,我们描述了与持久性细菌库形成、生存和肠道腔再播种相关的稀有事件(也称瓶颈)的定量分析。这为体内持久性细菌驱动的质粒转移提供了定量数据。虽然本章描述了在小鼠模型中分析沙门氏菌持久性细菌,但这些概念可以应用于任何实验系统,只要存在可追踪的实验系统即可。