Determination of bactericidal activity against 3HC-2-Tre-labelled Mycobacterium abscessus (Mycobacteroides abscessus) by automated fluorescence microscopy

IF 1.7 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS Journal of microbiological methods Pub Date : 2024-07-20 DOI:10.1016/j.mimet.2024.107002

Lea Mann, Fabienne Siersleben, Markus Lang, Adrian Richter

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Abstract

The minimum bactericidal concentration (MBC) of antibiotics is an important parameter for the potency of a drug in eradicating a bacterium as well as an important measure of the potential of a drug candidate in research and development. We have established a fluorescence-based microscopy method for the determination of MBCs against the non-tuberculous mycobacterium Mycobacterium abscessus (Mycobacteroides abscessus) to simplify and accelerate the performance of MBC determination compared to counting colony forming units on agar. Bacteria are labelled with the trehalose-coupled dye 3HC-2-Tre and analysed in a 96-well plate. The results of the new method are consistent with MBC determination by plating on agar. The method was used to evaluate the bactericidality of the antibiotics rifabutin, moxifloxacin, amikacin, clarithromycin and bedaquiline. Bactericidal effects against M. abscessus were observed, which are consistent with literature data.

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利用自动荧光显微镜测定对 3HC-2-Tre 标记的脓肿分枝杆菌（脓肿分枝杆菌）的杀菌活性。

抗生素的最小杀菌浓度（MBC）是衡量药物根除细菌效力的重要参数，也是衡量候选药物研发潜力的重要指标。我们建立了一种基于荧光的显微镜方法，用于测定对非结核分枝杆菌脓肿分枝杆菌（Mycobacteroides abscessus）的 MBC，与在琼脂上计数菌落形成单位相比，这种方法简化并加快了 MBC 的测定。细菌用三卤糖偶联染料 3HC-2-Tre 标记，并在 96 孔板中进行分析。新方法的结果与琼脂上的菌落形成单位测定法一致。该方法用于评估利福布丁、莫西沙星、阿米卡星、克拉霉素和贝达喹啉等抗生素的杀菌性。观察到了对脓肿霉菌的杀菌效果，这与文献数据一致。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.