Application of a Tetrazolium Salt with a Water-Soluble Formazan as an Indicator of Viability in Respiring Bacteria
Application of a Tetrazolium Salt with a Water-Soluble Formazan as an Indicator of Viability in Respiring Bacteria
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应用四唑盐与水溶性甲臜作为呼吸细菌活力的指示剂
DOI：
10.1128/aem.59.9.2891-2896.1993
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发表时间：
1993
影响因子：
4.4
通讯作者：
G. King
中科院分区：
文献类型：
--
作者：
P. Roslev;G. King
研究了四唑盐3′-{1-[（苯氨基）-羰基]-3，4-四唑}-双（4-甲氧基-6-硝基）苯磺酸钠水合物（XTT）作为呼吸细菌活力的比色指示剂。XTT被发孢甲基弯菌OB 3b、恶臭假单胞菌、大肠杆菌和枯草芽孢杆菌还原为橙子水溶性甲瓒产物。福尔马赞生产成比例的活细胞生物量，和XTT减少了所有的文化在没有添加电子偶联剂。通过M减少XTT。丝孢霉OB 3b在数小时内是线性的，并且通过外源底物（甲醇）的存在来刺激。在有氧条件下培养的培养物中加入氰化物，XTT减少量最初增加了10倍。在不存在外源性碳底物的情况下孵育的细菌的活力测量为XTT减少，并与来自平板计数的活力估计值进行比较。两种方法获得的结果基本相当，但当平板上的细胞回收率较低时，XTT测定法具有上级优势。E.大肠杆菌培养7天，在没有外源性碳底物的情况下，存活率降低90%，而M.发孢霉OB 3 B、恶臭假单胞菌和B。枯草芽孢杆菌的比例低于40%。
The tetrazolium salt sodium 3′-{1-[(phenylamino)-carbonyl]-3,4-tetrazolium}-bis (4-methoxy-6-nitro)benzene-sulfonic acid hydrate (XTT) was examined for use as a colorimetric indicator of viability in respiring bacteria. XTT was reduced to an orange, water-soluble formazan product by Methylosinus trichosporium OB3b, Pseudomonas putida, Escherichia coli, and Bacillus subtilis. Formazan production was proportional to live cell biomass, and XTT was reduced by all cultures in the absence of added electron-coupling agents. XTT reduction by M. trichosporium OB3b was linear over several hours and was stimulated by the presence of an exogenous substrate (methanol). Addition of cyanide to cultures incubated under oxic conditions gave an initial 10-fold increase in XTT reduction. Viability of bacteria incubated in the absence of exogenous carbon substrates was measured as XTT reduction and compared with viability estimates from plate counts. Results obtained with the two methods were generally comparable, but the XTT assay was superior when cell recovery on plates was low. Incubation of E. coli for 7 days in the absence of exogenous carbon substrates decreased viability by 90%, whereas the corresponding decreases for cultures of M. trichosporium OB3b, P. putida, and B. subtilis were less than 40%.