Hare Patricia J, LaGree Travis J, Byrd Brandon A, DeMarco Angela M, Mok Wendy W K
Department of Molecular Biology & Biophysics, UConn Health, Farmington, CT 06032, USA.
School of Dental Medicine, University of Connecticut, Farmington, CT 06032, USA.
Microorganisms. 2021 Nov 1;9(11):2277. doi: 10.3390/microorganisms9112277.
Antibiotic persistence is a phenomenon in which rare cells of a clonal bacterial population can survive antibiotic doses that kill their kin, even though the entire population is genetically susceptible. With antibiotic treatment failure on the rise, there is growing interest in understanding the molecular mechanisms underlying bacterial phenotypic heterogeneity and antibiotic persistence. However, elucidating these rare cell states can be technically challenging. The advent of single-cell techniques has enabled us to observe and quantitatively investigate individual cells in complex, phenotypically heterogeneous populations. In this review, we will discuss current technologies for studying persister phenotypes, including fluorescent tags and biosensors used to elucidate cellular processes; advances in flow cytometry, mass spectrometry, Raman spectroscopy, and microfluidics that contribute high-throughput and high-content information; and next-generation sequencing for powerful insights into genetic and transcriptomic programs. We will further discuss existing knowledge gaps, cutting-edge technologies that can address them, and how advances in single-cell microbiology can potentially improve infectious disease treatment outcomes.
抗生素耐受性是一种现象，即克隆细菌群体中的少数细胞能够在杀死其同类细胞的抗生素剂量下存活，尽管整个群体在基因上是敏感的。随着抗生素治疗失败率的上升，人们越来越有兴趣了解细菌表型异质性和抗生素耐受性背后的分子机制。然而，阐明这些稀有细胞状态在技术上可能具有挑战性。单细胞技术的出现使我们能够观察和定量研究复杂的、表型异质群体中的单个细胞。在这篇综述中，我们将讨论目前用于研究耐受性表型的技术，包括用于阐明细胞过程的荧光标签和生物传感器；流式细胞术、质谱、拉曼光谱和微流控技术的进展，这些技术能提供高通量和高内涵信息；以及用于深入了解遗传和转录组程序的新一代测序技术。我们还将进一步讨论现有的知识空白、能够解决这些空白的前沿技术，以及单细胞微生物学的进展如何有可能改善传染病的治疗效果。
