Tsukatani T, Suenaga H, Shiga M, Matsumoto K
Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, Kurume, Japan.
Lett Appl Microbiol. 2014 Aug;59(2):184-92. doi: 10.1111/lam.12264. Epub 2014 May 5.
A rapid microplate method for the proliferation assay of fungi and the antifungal susceptibility testing using the colorimetric microbial viability assay based on the reduction in a tetrazolium salt 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) with 2-methyl-1,4-napthoquinone as the electron mediator was developed. The proposed method was useful to measure the proliferation of 18 kinds of moulds and seven kinds of yeasts, including representative pathogens such as Aspergillus spp., Candida spp. and Cryptococcus spp. Linear relationships between the absorbance and viable fungal cell density were obtained for all fungi, suggesting that the absorbance change reflected the fungal proliferation. In addition, the minimum inhibitory concentrations (MICs) against a variety of different pathogenic moulds and yeasts for amphotericin B, itraconazole and 5-flucytosine were determined by susceptibility testing using the proposed method and compared with those obtained using the conventional broth microdilution method. There was an excellent agreement between the results obtained using the WST-8 colorimetric method and those obtained using the conventional Clinical and Laboratory Standard Institute method. The WST-8 colorimetric assay is a useful method for rapid determination of accurate MICs for a variety of different fungi.
A rapid microplate method for the proliferation assay of fungi and the antifungal susceptibility testing using the colorimetric microbial viability assay based on reduction in a tetrazolium salt (WST-8) was developed. The WST-8 colorimetric method was useful to measure the proliferation of a variety of different fungi. In the antifungal susceptibility testing, there was a good agreement between the MICs determined after 24 h using the WST-8 colorimetric method and those obtained after 48-96 h using the broth microdilution method. The proposed method was superior to conventional methods in terms of its rapidity towards a variety of different fungi.
开发了一种基于四唑盐2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯基)-2H-四唑单钠盐（WST-8）还原反应的比色微生物活力测定法的快速微孔板法，用于真菌增殖测定和抗真菌药敏试验，以2-甲基-1,4-萘醌作为电子介质。所提出的方法可用于测量18种霉菌和7种酵母的增殖，包括曲霉属、念珠菌属和隐球菌属等代表性病原体。所有真菌的吸光度与活真菌细胞密度之间均获得了线性关系，表明吸光度变化反映了真菌增殖。此外，通过所提出的方法进行药敏试验，测定了两性霉素B、伊曲康唑和5-氟胞嘧啶对多种不同致病霉菌和酵母的最低抑菌浓度（MIC），并与使用传统肉汤微量稀释法获得的结果进行了比较。使用WST-8比色法获得的结果与使用传统临床和实验室标准协会方法获得的结果之间具有极好的一致性。WST-8比色测定法是一种用于快速准确测定多种不同真菌MIC的有用方法。
开发了一种基于四唑盐（WST-8）还原反应的比色微生物活力测定法的快速微孔板法，用于真菌增殖测定和抗真菌药敏试验。WST-8比色法可用于测量多种不同真菌的增殖。在抗真菌药敏试验中，使用WST-8比色法在24小时后测定的MIC与使用肉汤微量稀释法在48-96小时后获得的MIC之间具有良好的一致性。所提出的方法在针对多种不同真菌的快速性方面优于传统方法。
