Bacterial biofilms demonstrate high broad-spectrum adaptive antibiotic resistance and cause two thirds of all infections, but there is a lack of approved antibiofilm agents. Unlike the standard minimal inhibitory concentration assay to assess antibacterial activity against planktonic cells, there is no standardized method to evaluate biofilm inhibition and/or eradication capacity of novel antibiofilm compounds. The protocol described here outlines simple and reproducible methods for assessing the biofilm inhibition and eradication capacities of novel antibiofilm agents against adherent bacterial biofilms grown in 96-well microtiter plates. It employs two inexpensive dyes: crystal violet to stain adhered biofilm biomass and 2,3,5-triphenyl tetrazolium chloride to quantify metabolism of the biofilm cells. The procedure is accessible to any laboratory with a plate reader, requires minimal technical expertise or training and takes 4 or 5 d to complete. Recommendations for how biofilm inhibition and eradication results should be interpreted and presented are also described.

细菌生物膜表现出高广谱适应性抗生素耐药性，并导致所有感染的三分之二，但缺乏经批准的抗生物膜剂。与评估对浮游细胞的抗菌活性的标准最小抑制浓度测定不同，没有标准化的方法来评估新型抗生物膜化合物的生物膜抑制和/或根除能力。此处描述的协议概述了用于评估新型抗生物膜剂对 96 孔微量滴定板中生长的粘附细菌生物膜的生物膜抑制和根除能力的简单且可重复的方法。它采用两种廉价染料：结晶紫染色粘附的生物膜生物量和 2,3,5-三苯基氯化四唑来量化生物膜细胞的新陈代谢。任何具有读板器的实验室都可以访问该程序，需要最少的技术专业知识或培训，并且需要 4 或 5 天才能完成。还描述了关于如何解释和呈现生物膜抑制和根除结果的建议。