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基于发光的曼氏血吸虫幼虫期药物筛选的低通量和中通量检测方法。

Luminescence-Based, Low- and Medium-Throughput Assays for Drug Screening in Schistosoma mansoni Larval Stage.

机构信息

National Research Council, Institute of Biochemistry and Cell Biology, Rome, Italy.

Translational Biology Department, IRBM S.p.A., Rome, Italy.

出版信息

Methods Mol Biol. 2020;2151:219-227. doi: 10.1007/978-1-0716-0635-3_18.

Abstract

Schistosomiasis is one of the major parasitic diseases with more than  200 million people infected worldwide every year. Praziquantel is the drug of choice against the schistosomiasis although the use of a single drug to treat such a large amount of infected people appears particularly worrisome. For this reason, the search of new schistosomicidal compounds is viewed as an urgent goal and a number of screening campaigns have been carried out in the past years. The larval stage of Schistosoma (schistosomula) has been widely used in order to identify new compounds against the parasite. Here we describe detailed practical procedures for a luminescence-based assay proven to be highly effective for the selection of schistosomicidal compounds on small and medium-high scale. The assay is based on the quantitation of the parasite ATP, a good indicator of metabolically active cells, as measure of schistosomula viability. This assay is fast and reproducible, and it is suitable either for manual or for semiautomated screenings.

摘要

血吸虫病是一种主要的寄生虫病,全球每年有超过 2 亿人感染。尽管使用单一药物治疗如此大量的感染者似乎特别令人担忧,但吡喹酮是治疗血吸虫病的首选药物。出于这个原因,寻找新的杀血吸虫化合物被视为当务之急,过去几年已经进行了多次筛选活动。血吸虫的幼虫期(尾蚴)已被广泛用于鉴定针对寄生虫的新化合物。在这里,我们描述了一种基于发光的检测方法的详细实用程序,该方法已被证明在小规模和中大规模筛选杀血吸虫化合物方面非常有效。该测定基于寄生虫 ATP 的定量,ATP 是代谢活跃细胞的良好指标,可作为尾蚴活力的衡量标准。该测定快速且可重复,既适用于手动筛选,也适用于半自动筛选。

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