微量滴定板中生长的微生物生物膜定量多种方法的比较
Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates.
作者信息
Peeters Elke, Nelis Hans J, Coenye Tom
机构信息
Laboratory for Pharmaceutical Microbiology, Ghent University, Harelbekestraat 72, Ghent, Belgium.
出版信息
J Microbiol Methods. 2008 Feb;72(2):157-65. doi: 10.1016/j.mimet.2007.11.010. Epub 2007 Nov 21.
In the present study six assays for the quantification of biofilms formed in 96-well microtiter plates were optimised and evaluated: the crystal violet (CV) assay, the Syto9 assay, the fluorescein diacetate (FDA) assay, the resazurin assay, the XTT assay and the dimethyl methylene blue (DMMB) assay. Pseudomonas aeruginosa, Burkholderia cenocepacia, Staphylococcus aureus, Propionibacterium acnes and Candida albicans were used as test organisms. In general, these assays showed a broad applicability and a high repeatability for most isolates. In addition, the estimated numbers of CFUs present in the biofilms show limited variations between the different assays. Nevertheless, our data show that some assays are less suitable for the quantification of biofilms of particular isolates (e.g. the CV assay for P. aeruginosa).
在本研究中，对用于定量96孔微量滴定板中形成的生物膜的六种测定方法进行了优化和评估：结晶紫（CV）测定法、Syto9测定法、荧光素二乙酸酯（FDA）测定法、刃天青测定法、XTT测定法和二甲基亚甲基蓝（DMMB）测定法。使用铜绿假单胞菌、洋葱伯克霍尔德菌、金黄色葡萄球菌、痤疮丙酸杆菌和白色念珠菌作为测试微生物。总体而言，这些测定方法对大多数分离株显示出广泛的适用性和高重复性。此外，生物膜中存在的估计菌落形成单位（CFU）数量在不同测定方法之间显示出有限的差异。然而，我们的数据表明，某些测定方法不太适合定量特定分离株的生物膜（例如，用于铜绿假单胞菌的CV测定法）。
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