Department of Pharmaceutical Microbiology and Bioanalysis, Medical University of Warsaw, Warsaw, Poland.
Department of Antibiotics and Microbiology, National Medicines Institute, Warsaw, Poland.
Antimicrob Agents Chemother. 2023 May 17;67(5):e0137322. doi: 10.1128/aac.01373-22. Epub 2023 Apr 6.
The analysis of antimicrobial activity is usually MIC- and minimal bactericidal concentration (MBC)-focused, though also crucial are resistance-related parameters, e.g., the frequency of spontaneous mutant selection (FSMS), the mutant prevention concentration (MPC), and the mutant selection window (MSW). -determined MPCs, however, are sometimes variable, poorly repeatable, and not always reproducible . We propose a new approach to the determination of MSWs, along with novel parameters: MPC-D, MSW-D (for dominant mutants, i.e., selected with a high frequency, without a fitness loss), and MPC-F, MSW-F (for inferior mutants, i.e., with an impaired fitness). We also propose a new method for preparing the high-density inoculum (>10 CFU/mL). In this study, the MPC and MPC-D (limited by FSMS of <10) of ciprofloxacin, linezolid, and novel benzosiloxaborole (No37) were determined for Staphylococcus aureus ATCC 29213 using the standard agar method, while the MPC-D and MPC-F were determined by the novel broth method. Regardless of the method, MSWs of linezolid and No37 were the same. However, MSWs of ciprofloxacin in the broth method was narrower than in the agar method. In the broth method, the 24-h incubation of ~10 CFU in a drug-containing broth differentiates the mutants that can dominate the cell population from those that can only be selected under exposure. We consider MPC-Ds in the agar method to be less variable and more repeatable than MPCs. Meanwhile, the broth method may decrease discrepancies between and MSWs. The proposed approaches may help establish MPC-D-related resistance-restricting therapies.
抗微生物活性的分析通常集中在 MIC 和最小杀菌浓度 (MBC) 上,尽管与耐药性相关的参数也很关键,例如自发突变选择频率 (FSMS)、突变预防浓度 (MPC) 和突变选择窗 (MSW)。然而,确定的 MPC 有时是可变的、重复性差的,并且并不总是可重现的。我们提出了一种新的方法来确定 MSWs,以及新的参数:MPC-D、MSW-D(用于优势突变体,即高频率选择,没有适应性损失)和 MPC-F、MSW-F(用于劣势突变体,即适应性受损)。我们还提出了一种新的高密度接种物制备方法(>10 CFU/mL)。在这项研究中,使用标准琼脂方法测定了金黄色葡萄球菌 ATCC 29213 中环丙沙星、利奈唑胺和新型苯并硅氧硼唑 (No37) 的 MPC 和 MPC-D(受 FSMS <10 的限制),而通过新的肉汤方法测定了 MPC-D 和 MPC-F。无论采用哪种方法,利奈唑胺和 No37 的 MSWs 都是相同的。然而,肉汤法中环丙沙星的 MSWs 比琼脂法窄。在肉汤方法中,含有药物的肉汤中 24 小时孵育可以区分能够主导细胞群体的突变体和只能在暴露下选择的突变体。我们认为琼脂方法中的 MPC-Ds 比 MPCs 更具可变性和重复性。同时,肉汤法可能会减少 和 MSWs 之间的差异。所提出的方法可能有助于建立与 MPC-D 相关的耐药限制治疗方法。