A RAPID COLORIMETRIC ASSAY OF FUNGAL VIABILITY WITH THE TETRAZOLIUM SALT MTT
A RAPID COLORIMETRIC ASSAY OF FUNGAL VIABILITY WITH THE TETRAZOLIUM SALT MTT
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DOI：
10.1093/infdis/152.5.938
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发表时间：
1985-01-01
影响因子：
6.4
通讯作者：
DIAMOND, RD
中科院分区：
文献类型：
--
作者：
LEVITZ, SM;DIAMOND, RD
描述了一种使用四唑盐四甲基偶氮唑盐的快速比色分析真菌活性的方法。所有被测试的各种医学上重要的真菌的活物种都降低了MTT值，结果在几个小时内形成了清晰可见的紫色结晶的MTTFormazan。该方法特别适用于测定99%的烟曲霉菌和米根霉菌丝在与四甲基偶氮唑盐孵育时染成紫色的单个菌丝的活力，而用任何一种方法杀死的菌丝的活力均为0。比较了用不同浓度的过氧化氢杀灭烟曲霉菌丝时，测定真菌活力的四甲基偶氮唑盐比色法和玻片培养法。用乙醇可以很容易地从真菌中提取出四甲基偶氮唑蓝-福尔马赞，这种方法可以用分光光度法定量测定无细胞和细胞系统中的真菌杀伤率。白念珠菌芽胞和烟曲霉菌丝等几种真菌的MTT值与接种量呈线性关系。用四甲基偶氮唑盐比色法测定人白细胞对白色念珠菌芽胞的杀灭率为80.3%，而用稀释法和倒置平板法测定的杀灭率为70.5%。用四甲基偶氮唑盐比色法测定了人白细胞对烟曲霉菌丝的杀灭作用。
A rapid colorimetric assay for viability of fungi that uses the tetrazolium salt MTT is described. All live species of a wide range of medically important fungi that were tested reduced MTT, with resultant intracellular formation within several hours of clearly visible purple crystals of MTT-formazan. The assay proved particularly useful in measuring viability of individual hyphae; > 99% of live Aspergillus fumigatus and Rhizopus oryzae hyphae stained purple when incubated with MTT compared with 0% of hyphae killed by any of a variety of methods. The MTT method for measuring fungal viability closely correlated with a slide-culture technique when killing of A. fumigatus hyphae by graded concentrations of hydrogen peroxide was compared. MTT-formazan was easily extracted from the fungi by alcohol, a procedure allowing spectrophotometric quantification of fungal killing in both cell-free and cellular systems. A linear relation between inocula and MTT reduction of several fungi, including Candida albicans blastospores and A. fumigatus hyphae, was demonstrated. Human leukocytes killed 80.3% of C. albicans blastospores as measured by the MTT assay, compared with 70.5% as measured by dilutions and pour plates. Killing of A. fumigatus hyphae by human leukocytes was also quantitated by using the MTT assay.