NOVEL METHODS FOR RAPID DRUG SUSCEPTIBILITY TESTING AND DRUG SCREENS FOR GROWING AND NON-GROWING CELLS OF PROKARYOTIC AND EUKARYOTIC ORIGIN
申请人:Zhang, Ying
公开号:EP3321371A1
公开(公告)日:2018-05-16
The presently disclosed subject matter provides methods, compositions, and kits for extremely rapid assessment of drug susceptibility of growing as well as non-growing cells of prokaryotic and also eukaryotic origins, without the need for organism growth, which is in contrast to the current growth-dependent methods.This growth-independent new methodology achieves extremely rapid drug susceptibility testing results previously not possible. Methods for rapidly identifying compounds with inhibitory activity against both growing and non-growing cells in real-time continuous manner with previously unachievable efficiency in drug screens using the novel methodology are also provided. Cell cultures are incubated with a drug or test agent and with a staining mixture containing a first fluorescent dye indicative the emission of which is indicative of live cells (e.g. SYBR Green I) and a second fluorescent dye indicative emitting a different color, the emission of which is indicative of dead cells (e.g. Propidium iodide). The ratio of the intensities of the emitted fluorescence of the dyes is indicative of the percentage live and dead cells in the culture.
与目前依赖生长的方法相比,本公开的主题提供了无需生物体生长即可对原核和真核的生长和非生长细胞的药物敏感性进行极快速评估的方法、组合物和试剂盒,这种不依赖生长的新方法实现了以前无法实现的极快速药物敏感性测试结果。此外,还提供了实时连续地快速鉴定对生长细胞和非生长细胞具有抑制活性的化合物的方法,这种方法在使用新方法进行药物筛选时具有以前无法实现的效率。细胞培养物与药物或测试剂以及染色混合物一起孵育,染色混合物含有第一种荧光染料指示剂,其发射的荧光指示剂可指示活细胞(如 SYBR 绿 I)和第二种发出不同颜色的荧光染料指示剂,其发射的荧光指示剂可指示死细胞(如碘化丙啶)。染料发射荧光强度的比值表示培养液中活细胞和死细胞的百分比。