Shimony Orly, Jaffe Charles L
The Kuvin Centre for the Study of Infectious and Tropical Diseases, Hebrew University - Hadassah Medical School, P.O. Box 12272, Jerusalem 91220, Israel.
J Microbiol Methods. 2008 Oct;75(2):196-200. doi: 10.1016/j.mimet.2008.05.026. Epub 2008 Jun 5.
A rapid fluorescent viability assay employing alamarBlue was optimized for use with Leishmania axenic amastigotes, the stage of the parasite responsible for disease pathology. The activity of two protein kinase inhibitors, Staurosporine and H-89, as well as Amphotericin B, on promastigotes and amastigotes of Leishmania donovani and Leishmania tropica was compared. Both protein kinase inhibitors inhibited promastigote growth at lower concentrations than amastigotes, while the GI(50) for Amphotericin B on both stages was similar. This assay only requires a limited number of axenic amastigotes (50,000 cells/well) and can be used to rapidly screen large chemical or natural product libraries for activity against amastigotes.
一种使用alamarBlue的快速荧光活力测定法已针对利什曼原虫无鞭毛体（该寄生虫导致疾病病理的阶段）进行了优化。比较了两种蛋白激酶抑制剂（星形孢菌素和H - 89）以及两性霉素B对杜氏利什曼原虫和热带利什曼原虫前鞭毛体和无鞭毛体的活性。两种蛋白激酶抑制剂在比无鞭毛体更低的浓度下就能抑制前鞭毛体的生长，而两性霉素B在两个阶段的半数生长抑制浓度（GI(50)）相似。该测定法仅需要有限数量的无鞭毛体（50,000个细胞/孔），并且可用于快速筛选大型化学或天然产物文库，以寻找针对无鞭毛体的活性物质。
