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可溶珠:一种用于评估抗菌耐药性的新型体外生物膜模型。

The dissolvable bead: A novel in vitro biofilm model for evaluating antimicrobial resistance.

作者信息

Dall G F, Tsang S T J, Gwynne P J, Wilkinson A J, Simpson A H R W, Breusch S J B, Gallagher M P

机构信息

School of Biological Sciences, University of Edinburgh, Darwin Building, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, United Kingdom; Department of Orthopaedic surgery, University of Edinburgh, Chancellor's building, 49 Little France Crescent, Old Dalkeith Road, Edinburgh EH16 4SB, United Kingdom.

School of Biological Sciences, University of Edinburgh, Darwin Building, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, United Kingdom; Department of Orthopaedic surgery, University of Edinburgh, Chancellor's building, 49 Little France Crescent, Old Dalkeith Road, Edinburgh EH16 4SB, United Kingdom; Department of Orthopaedic surgery, Royal Infirmary of Edinburgh, 51 Little France, Old Dalkeith Road, Edinburgh EH16 4SA, United Kingdom.

出版信息

J Microbiol Methods. 2017 Nov;142:46-51. doi: 10.1016/j.mimet.2017.08.020. Epub 2017 Sep 17.

DOI:10.1016/j.mimet.2017.08.020
PMID:28870772
Abstract

In vitro biofilm assays are a vital first step in the assessment of therapeutic effectiveness. Current biofilm models have been found to be limited by throughput, reproducibility, and cost. We present a novel in vitro biofilm model, utilising a sodium alginate substratum for surface biofilm colony formation, which can be readily dissolved for accurate evaluation of viable organisms. The dissolving bead biofilm assay was evaluated using a range of clinically relevant strains. The reproducibility and responsiveness of the assay to an antimicrobial challenge was assessed using standardised methods. Cryo-scanning electron microscopy was used to image biofilm colonies. Biofilms were grown for 20h prior to testing. The model provides a reproducible and responsive assay to clinically-relevant antimicrobial challenges, as defined by established guidelines. Moreover cryo-scanning electron microscopy demonstrates that biofilm formation is localised exclusively to the alginate bead surface. Our results suggest that this simple model provides a robust and adaptable assay for the investigation of bacterial biofilms.

摘要

体外生物膜检测是评估治疗效果至关重要的第一步。目前已发现现有的生物膜模型在通量、可重复性和成本方面存在局限性。我们提出了一种新型的体外生物膜模型,利用海藻酸钠基质进行表面生物膜菌落形成,该基质可轻松溶解以准确评估活菌数量。使用一系列临床相关菌株对溶解珠生物膜检测进行了评估。采用标准化方法评估了该检测对抗菌挑战的可重复性和反应性。利用冷冻扫描电子显微镜对生物膜菌落进行成像。在测试前,生物膜培养20小时。该模型针对既定指南所定义的临床相关抗菌挑战提供了一种可重复且有反应的检测方法。此外,冷冻扫描电子显微镜显示生物膜形成仅局限于海藻酸钠珠表面。我们的结果表明,这个简单的模型为细菌生物膜的研究提供了一种强大且适应性强的检测方法。

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