Lin Jing-Wen, Sajid Mohammed, Ramesar Jai, Khan Shahid M, Janse Chris J, Franke-Fayard Blandine
Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
Methods Mol Biol. 2013;923:507-22. doi: 10.1007/978-1-62703-026-7_35.
We describe two improved assays for in vitro and in vivo screening of inhibitors and chemicals for antimalarial activity against blood stages of the rodent malaria parasite, Plasmodium berghei. These assays are based on the determination of bioluminescence in small blood samples that is produced by reporter parasites expressing luciferase. Luciferase production increases as the parasite develops in a red blood cell and as the numbers of parasites increase during an infection. In the first assay, in vitro drug luminescence (ITDL) assay, the in vitro development of ring-stage parasites into mature schizonts in the presence and absence of candidate inhibitor(s) is quantified by measuring luciferase activity after the parasites have been allowed to mature into schizonts in culture. In the second assay, the in vivo drug luminescence (IVDL) assay, in vivo parasite growth (using a standard 4-day suppressive drug test) is quantified by measuring the luciferase activity of circulating parasites in samples of tail blood of drug-treated mice.
我们描述了两种经过改进的检测方法，用于体外和体内筛选针对啮齿动物疟原虫伯氏疟原虫血液阶段具有抗疟活性的抑制剂和化学物质。这些检测方法基于对表达荧光素酶的报告寄生虫在小血样中产生的生物发光的测定。随着寄生虫在红细胞中发育以及感染过程中寄生虫数量的增加，荧光素酶的产生也会增加。在第一种检测方法，即体外药物发光（ITDL）检测中，通过在培养中使寄生虫发育成裂殖体后测量荧光素酶活性，来量化环状阶段寄生虫在有或没有候选抑制剂存在的情况下体外发育成成熟裂殖体的情况。在第二种检测方法，即体内药物发光（IVDL）检测中，通过测量经药物处理的小鼠尾血样本中循环寄生虫的荧光素酶活性，来量化体内寄生虫的生长情况（使用标准的4天抑制性药物试验）。
