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黏菌素在标准实验条件下大量丢失。

Colistin Is Extensively Lost during Standard Experimental Conditions.

机构信息

Department of Medical Sciences, Uppsala University, Uppsala, Sweden

Department of Medical Sciences, Uppsala University, Uppsala, Sweden.

出版信息

Antimicrob Agents Chemother. 2017 Oct 24;61(11). doi: 10.1128/AAC.00857-17. Print 2017 Nov.

Abstract

Colistin adheres to a range of materials, including plastics in labware. The loss caused by adhesion influences an array of methods detrimentally, including MIC assays and time-kill experiments. The aim of this study was to characterize the extent and time course of colistin loss in different types of laboratory materials during a simulated time-kill experiment without bacteria or plasma proteins present. Three types of commonly used large test tubes, i.e., soda-lime glass, polypropylene, and polystyrene, were studied, as well as two different polystyrene microplates and low-protein-binding microtubes. The tested concentration range was 0.125 to 8 mg/liter colistin base. Exponential one-phase and two-phase functions were fitted to the data, and the adsorption of colistin to the materials was modeled with the Langmuir adsorption model. In the large test tubes, the measured start concentrations ranged between 44 and 102% of the expected values, and after 24 h, the concentrations ranged between 8 and 90%. The half-lives of colistin loss were 0.9 to 12 h. The maximum binding capacities of the three materials ranged between 0.4 and 1.1 μg/cm, and the equilibrium constants ranged between 0.10 and 0.54 ml/μg. The low-protein-binding microtubes showed start concentrations between 63 and 99% and concentrations at 24 h of between 59 and 90%. In one of the microplates, the start concentrations were below the lower limit of quantification at worst. In conclusion, to minimize the effect of colistin loss due to adsorption, our study indicates that low-protein-binding polypropylene should be used when possible for measuring colistin concentrations in experimental settings, and the results discourage the use of polystyrene. Furthermore, when diluting colistin in protein-free media, the number of dilution steps should be minimized.

摘要

黏菌素会黏附于多种材料,包括实验器皿中的塑料。黏附造成的损失会对多种方法产生负面影响,包括 MIC 测定和杀菌动力学实验。本研究旨在模拟无细菌和血浆蛋白存在的杀菌动力学实验,对不同实验室材料中黏菌素的损失程度和时间进程进行特征描述。研究中使用了三种常用的大型测试管,即钠钙玻璃、聚丙烯和聚苯乙烯,以及两种不同的聚苯乙烯微孔板和低蛋白结合微管。测试浓度范围为 0.125 至 8 mg/L 黏菌素碱。采用指数一相和两相函数对数据进行拟合,并采用朗缪尔吸附模型对黏菌素在材料上的吸附进行建模。在大型测试管中,测量的起始浓度介于预期值的 44%至 102%之间,24 小时后浓度介于 8%至 90%之间。黏菌素损失的半衰期为 0.9 至 12 小时。三种材料的最大结合容量介于 0.4 至 1.1 μg/cm,平衡常数介于 0.10 至 0.54 ml/μg。低蛋白结合微管的起始浓度介于 63%至 99%之间,24 小时后的浓度介于 59%至 90%之间。在其中一个微孔板中,起始浓度最差时低于定量下限。总之,为了最大程度地减少因吸附造成的黏菌素损失的影响,我们的研究表明,在实验环境中测量黏菌素浓度时,应尽可能使用低蛋白结合的聚丙烯,同时不鼓励使用聚苯乙烯。此外,在无蛋白介质中稀释黏菌素时,应尽量减少稀释步骤的数量。

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Colistin Is Extensively Lost during Standard Experimental Conditions.黏菌素在标准实验条件下大量丢失。
Antimicrob Agents Chemother. 2017 Oct 24;61(11). doi: 10.1128/AAC.00857-17. Print 2017 Nov.

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