Division of Inflammation and Infection, Lab 1, floor 12, Department of Biomedical and Clinical Sciences, Faculty of Medicine and Health Sciences, Linköping University, SE-58185, Linköping, Sweden.
Division of Clinical Microbiology, Department of Biomedical and Clinical Sciences, Faculty of Medicine and Health Sciences, Linköping University, SE-58185, Linköping, Sweden.
BMC Microbiol. 2021 Jun 5;21(1):167. doi: 10.1186/s12866-021-02212-3.
Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs.
Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype.
Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.
高效高通量药物筛选测定法对于发现新的抗分枝杆菌药物是必要的。我们的工作目的是开发和验证一种基于活细胞成像的测定法，该方法可监测两种不同表型的结核分枝杆菌的生长，并测试它们对常用结核病药物的敏感性。
使用活细胞成像系统 IncuCyte S3 成功地将浮游和缠结表型作为荧光物体进行监测，从而收集描述聚集体大小和生长的不同特征的数据。对聚集体总面积变化的定量分析用于定义选定结核病药物的 IC 和 MIC 值，结果表明缠结表型生长更快，对利福平的敏感性更高。棋盘式方法测试亚抑制浓度药物的配对组合，利福平、利奈唑胺和普托马尼德显示对缠结表型具有更好的生长抑制作用。
我们的结果强调了使用自动化活细胞成像及其在高通量全细胞筛选中的潜力，以评估现有和寻找新型抗分枝杆菌药物的效率。
