Woehrmann Marcos H, Gassner Nadine C, Bray Walter M, Stuart Joshua M, Lokey Scott
Department of Biomolecular Engineering, UC Santa Cruz, Santa Cruz, California, USA.
J Biomol Screen. 2010 Feb;15(2):196-205. doi: 10.1177/1087057109355060. Epub 2010 Jan 19.
A high-throughput (HT) agar-based halo assay is described, which allows for rapid screening of chemical libraries for bioactivity in microorganisms such as yeast and bacteria. A pattern recognition algorithm was developed to identify halo-like shapes in plate reader optical density (OD) measurements. The authors find that the total growth inhibition within a detected halo provides an accurate estimate of a compound's potency measured in terms of its EC(50). The new halo recognition method performs significantly better than an earlier method based on single-point OD readings. An assay based on the halo algorithm was used to screen a 21,120-member library of drug-like compounds in Saccharomyces cerevisiae, leading to the identification of novel bioactive scaffolds containing derivatives of varying potencies. The authors also show that the HT halo assay can be performed with the pathogenic bacterium Vibrio cholerae and that liquid culture EC(50) values and halo scores show a good correlation in this organism. These results suggest that the HT halo assay provides a rapid and inexpensive way to screen for bioactivity in multiple microorganisms.
本文描述了一种基于琼脂的高通量（HT）光晕测定法，该方法可用于快速筛选化学文库，以检测酵母和细菌等微生物中的生物活性。开发了一种模式识别算法，用于在酶标仪光密度（OD）测量中识别类似光晕的形状。作者发现，检测到的光晕内的总生长抑制率能够准确估计化合物以其半数有效浓度（EC50）衡量的效力。新的光晕识别方法比基于单点OD读数的早期方法表现显著更好。基于光晕算法的测定法用于在酿酒酵母中筛选一个包含21120种类药物化合物的文库，从而鉴定出含有不同效力衍生物的新型生物活性支架。作者还表明，HT光晕测定法可用于致病性细菌霍乱弧菌，并且该方法在该生物体中测得的液体培养EC50值与光晕评分显示出良好的相关性。这些结果表明，HT光晕测定法提供了一种快速且廉价的方法来筛选多种微生物中的生物活性。
