用于曲霉菌种抗真菌药敏试验的比色测定法。
Colorimetric assay for antifungal susceptibility testing of Aspergillus species.
作者信息
Meletiadis J, Mouton J W, Meis J F, Bouman B A, Donnelly J P, Verweij P E
机构信息
Department of Medical Microbiology, University Medical Center Nijmegen, Nijmegen, The Netherlands.
出版信息
J Clin Microbiol. 2001 Sep;39(9):3402-8. doi: 10.1128/JCM.39.9.3402-3408.2001.
Abstract
A colorimetric assay for antifungal susceptibility testing of Aspergillus species (Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Aspergillus nidulans, and Aspergillus ustus) is described based on the reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-[(sulphenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) in the presence of menadione as an electron-coupling agent. The combination of 200 microg of XTT/ml with 25 microM menadione resulted in a high production of formazan within 2 h of exposure, allowing the detection of hyphae formed by low inocula of 10(2) CFU/ml after 24 h of incubation. Under these settings, the formazan production correlated linearly with the fungal biomass and less-variable concentration effect curves for amphotericin B and itraconazole were obtained.
摘要
本文描述了一种用于曲霉属菌种（烟曲霉、黄曲霉、土曲霉、构巢曲霉和黑曲霉）抗真菌药敏试验的比色测定法，该方法基于在甲萘醌作为电子偶联剂存在的情况下，四唑盐2,3-双(2-甲氧基-4-硝基-5-[(硫代苯氨基)羰基]-2H-四唑鎓-氢氧化物（XTT）的还原反应。每毫升200微克XTT与25微摩尔甲萘醌的组合在暴露2小时内导致大量甲臜生成，使得在孵育24小时后能够检测到由每毫升10(2) CFU的低接种量形成的菌丝。在这些条件下，甲臜生成与真菌生物量呈线性相关，并且获得了两性霉素B和伊曲康唑的浓度效应曲线，其变异性较小。
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