Comparison of three assays for the quantification of Candida biomass in suspension and CDC reactor grown biofilms
Comparison of three assays for the quantification of Candida biomass in suspension and CDC reactor grown biofilms
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DOI：
10.1016/j.mimet.2005.03.014
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发表时间：
2005-12-01
影响因子：
2.2
通讯作者：
Nelis, HJ
中科院分区：
文献类型：
--
作者：
Honraet, K;Goetghebeur, E;Nelis, HJ
测量生物膜中酵母代谢活性的常用测定法是基于四唑盐XTT {2，3-双（2-甲氧基-4-硝基-5-磺基苯基）-5-[（苯基氨基）羰基]-2H-氢氧化四唑}还原成有色甲瓒。然而，最近的一份报告，也证实了我们自己的研究结果的显色XTT测定的缺点，促使我们研究酵母生物量定量的替代方法。为此，对使用荧光素二乙酸酯（FDA）和SYTO（R）9的两种荧光测定法以及XTT测定法在白色念珠菌和近平滑念珠菌细胞数响应曲线的线性范围、精密度以及种内和种间变异性方面进行了比较评价。在多标记微量滴定板读数器中进行荧光和吸光度的阅读。所有三种测定法都足以测定嗜热酵母生物量，但FDA和SYTO（R）9测定法具有实际优势。当应用于CDC生物膜反应器中获得的酵母生物膜生物量的定量时，FDA测定被证明是上级的。(c)2005 Elsevier B. V.保留所有权利。
A common assay to measure yeast metabolic activity in biofilns is based on the reduction of the tetrazolium salt XTT {2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide} to a colored formazan. However, a recent report, also confirmed by our own findings about the shortcomings of the chromogenic XTT assay, has prompted us to investigate alternative methods for yeast biomass quantification. To this end, two fluorogenic assays using fluorescein diacetate (FDA) and SYTO (R) 9 as well as the XTT assay were comparatively evaluated with regard to the linear range of Candida albicans and Candida parapsilosis cell number-response curves, precision and intra- and interspecies variability. Reading of fluorescence and absorbance was carried out in a multilabel microtiter plate reader. All three assays were adequate for the determination of planktonic yeast biomass, but the FDA and SYTO (R) 9 assays present practical advantages. When applied to the quantification of yeast biofilm biomass obtained in the CDC biofilm reactor, the FDA assay proved superior. (c) 2005 Elsevier B.V. All rights reserved.