School of Life and Health Sciences, Aston University, Birmingham, UK.
Methods Mol Biol. 2021;2314:247-260. doi: 10.1007/978-1-0716-1460-0_10.
Non-replicating persistence (NRP) is a functional adaptation that mycobacteria undergo in response to the stresses of the granuloma, facilitating antibiotic tolerance and long-term infection. These stresses, or NRP-inducing factors, include hypoxia, nutrient deprivation, and nitric oxide assault, which mycobacteria are well evolved to tolerate through a series of metabolic and physiological adaptations producing the NRP state. Most attempts to replicate these conditions in vitro have focused on only one of these factors at a time for ease and simplicity, but as a result, do not necessarily produce physiologically relevant phenotypes. Here, we provide the methods for two different in vitro NRP strategies that are useful for drug susceptibility testing and high-throughput screening.
非复制性持续存在（NRP）是分枝杆菌对肉芽肿压力的一种功能适应，有助于抗生素耐受和长期感染。这些压力或 NRP 诱导因子包括缺氧、营养缺乏和一氧化氮攻击，分枝杆菌通过一系列代谢和生理适应很好地耐受这些压力，产生 NRP 状态。大多数试图在体外复制这些条件的尝试都侧重于一次只关注这些因素中的一个，以方便和简化，但结果并不一定产生生理相关的表型。在这里，我们提供了两种不同的体外 NRP 策略的方法，这些方法对药物敏感性测试和高通量筛选很有用。
