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多孔板-MTT比色法测定植物抗菌成分对细菌的抑制活性 预览 被引量:20

Detection of Plant Antibacterial Components by Using Microplate-MTT Colorimetric Assay
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摘要 多孔板-MTT比色法测定植物抗菌成分对细菌抑制活性的步骤为:每孔加入浓度为10^6cfu/mL的供试菌液90灿,然后加入不同浓度的药液10μL,28℃暗培养24h后,每孔中加入5mg/mL的MTT溶液10μL,继续培养4h后加入10%二甲基亚砜100μL,振荡30min,在570nm处测定溶液的吸光值。采用以上方法,测定植物抗菌成分蓝桉醇对辣椒斑点病黄单胞菌(Xanthomonas vesicatoria)和枯草芽孢杆菌(Bacillus subtilis)的半抑制浓度(IC50)分别为0.158和0.395mg/mL,小檗碱对溶血葡萄球菌(Staphylococcus haemolyticus)的IC50为0.587mg/mL。结果表明,多孔板-MTT比色法能快速、微量地测定植物成分对细菌的抑制活性。 An optimal experimental procedure with microplate-MTT colorimetric assay was obtained ,that was : 1 ) addition of 90 μL of bacterial suspension in concentration of 106 cfu/mL in each well of a 96-well micro plate ;2) then addition of 10 μL of the component solution with different concentration ;3 ) cultivation for 24 h at 28 0℃ in darkness ;4) then 10 μL of MTT solution in 5 mg/mL was added in each well;5) after incubation for another 4 h at 28 ℃ ,100 μL of 10% DMSO was added;6) after shaking for another 30 min ,light absorption values were obtained at 570 nm for the snpernatant with a micro plate spectrophotometer. As the optimal MTT assay mentioned above, globulol from Eucalyptus globulus fruits showed strong inhibitory activities against Xanthomonas vesicatoria and Bacillus subtilis that median effective inhib- itory concentration( IC50 ) were 0. 158 and 0. 395 mg/mL respectively. Berberine from Phellodendron chinertse fruits had its strong inhibitory activity against Staphylococcus haemolyticus obtaining 0. 587 mg/mL of IC50. This indicated that plant antibacterial components could be evaluated fast and micro-quantitatively by using microplate-MTT colorimetric assay.
作者 唐静 谈满良 赵江林 汪冶 周立刚 TANG Jing,TAN Man-liang,ZHAO Jiang-lin , WANG Ye, ZHOU Li-gang (1. College of Agronomy and Bioteehnology, China Agricultural University,Beijing 100193, China ; 2. The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Seieuces , Guiyang 550002, China)
出处 《天然产物研究与开发》 CAS 2008年第6期 949-952,共4页 Natural Product Research and Development
基金 高等学校博士学科点专项科研基金(20060019048) 教育部新世纪优秀人才支持计划(NCET-05-0134) 国家自然科学基金(30871662)
关键词 多孔板-MTT比色法 抗菌活性成分 抗细菌活性 microplate-MTT colorimetric assay antimierobial components antibacterial activity
作者简介 通讯作者Tel:86-10-62731199;E-mail:lgzhou@cau.edu.cn
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参考文献11

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