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    • 6. 发明公开
    • NOVEL METHODS FOR RAPID DRUG SUSCEPTIBILITY TESTING AND DRUG SCREENS FOR GROWING AND NON-GROWING CELLS OF PROKARYOTIC AND EUKARYOTIC ORIGIN
    • 用于快速药物敏感性试验和药物屏障的新方法用于生长和非生长的原核和真核生成细胞
    • EP3321371A1
    • 2018-05-16
    • EP17159022.7
    • 2017-03-02
    • Zhang, YingFeng, JieYee, RebeccaZhang, ShuoZhang, WenhongWang, Xiaodong
    • ZHANG, YingFENG, JieYEE, RebeccaZHANG, WenhongZHANG, Shuo
    • C12Q1/18G01N33/50
    • G01N33/5014C12Q1/18Y02A50/57
    • 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.
    • 本发明公开的主题提供了方法,组合物和试剂盒,用于非常快速评估原核生物和真核生物起源的生长以及非生长细胞的药物敏感性,而不需要生物体生长,这与目前的生长相反 这种独立于生长的新方法实现了以前不可能的极快的药物敏感性测试结果。 还提供了用实时连续方式迅速鉴定具有抑制生长和非生长细胞的活性的化合物的方法,以及使用新方法在药物筛选中以前无法实现的效率。 将细胞培养物与药物或测试试剂以及含有第一荧光染料和第二荧光染料的染色混合物孵育,所述第一荧光染料指示其发射指示活细胞(例如SYBR Green I),第二荧光染料指示发射不同颜色, 这是死细胞的指示(例如碘化丙锭)。 染料荧光发射强度的比例表明培养物中活细胞和死细胞的百分比。