Development of a robust microliter plate-based assay method for assessment of bioactivity
Development of a robust microliter plate-based assay method for assessment of bioactivity
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DOI：
10.1016/j.mimet.2004.04.017
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发表时间：
2004-09-01
影响因子：
2.2
通讯作者：
O'Shea, DG
中科院分区：
文献类型：
--
作者：
Casey, JT;O'Cleirigh, C;O'Shea, DG
开发了一种基于微量滴定板的测定法，用于定量监测吸水链霉菌发酵样品中生物活性化合物的产生。报告的方法证明了基于纸片扩散理论的生物活性评估方法成功应用于高通量分析的微量滴定测定。该测定方法有助于生成测试生物体（大肠杆菌、枯草芽孢杆菌和酿酒酵母）对生物活性化合物的剂量反应曲线。使用该剂量反应曲线，该方法有助于定义三个不同的最低抑制浓度 (MIC) 值，用于表征样品的生物活性属性。该测定使用既定的标准程序，以方便该测定适应更广泛的测试微生物。由于加入了将浊度转换为生物量浓度的步骤，用于计算生物活性，因此由于假设浊度和生物量浓度之间的线性关系而产生的误差也减少了。 (C) 2004 Elsevier B.V. 保留所有权利。
A microtiter plate-based assay was developed for the quantitative monitoring of bioactive compound production in Streptomyces hygroscopicus fermentation samples. The method reported demonstrates the successful application of the theories of disk diffusion based methods of bioactivity assessment, to a microtiter assay for high throughput analysis. The assay method facilitates the generation of the dose-response curve of test organisms (Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae) to a bioactive compound. Using this dose-response curve, the method facilitates definition of three distinct Minimum Inhibitory Concentration (MIC) values for use in the characterisation of the bioactive attributes of a sample. The assay uses established standard procedures to facilitate adaptation of the assay for use with a wider range of test microorganisms. Errors due to the assumption of a linear relationship between turbidity and biomass concentration are also reduced, due to incorporation of a step to convert turbidity to biomass concentration, for use in the calculation of bioactivity. (C) 2004 Elsevier B.V. All rights reserved.