Luciferase-Based, High-Throughput Assay for Screening and Profiling Transmission-Blocking Compounds against Plasmodium falciparum Gametocytes
Luciferase-Based, High-Throughput Assay for Screening and Profiling Transmission-Blocking Compounds against Plasmodium falciparum Gametocytes
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DOI：
10.1128/aac.01949-15
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发表时间：
2016-04-01
影响因子：
4.9
通讯作者：
Avery, Vicky M.
中科院分区：
文献类型：
--
作者：
Lucantoni, Leonardo;Fidock, David A.;Avery, Vicky M.
发现能够同时针对恶性疟原虫的无性和配子体阶段的新的抗疟药物，对于消灭疟疾运动的成功至关重要。我们已经开发并验证了一个强大的，快速的，具有成本效益的高通量报告基因检测，以确定化合物对晚期（阶段IV和V）配子体的活性。该测定法适用于在长达72小时的孵育时间下测试化合物活性，表现出优异的质量和再现性，平均Z'值为0.85 +/-0.01。我们使用该方法从三个化学成分不同的库中筛选了10，000多种化合物。筛选结果强调了使用已知对寄生虫无性阶段具有活性的化合物集合作为杀配子活性检测的起点以最大化鉴定杀配子化合物的机会的机会。该测定扩展了我们先前报道的荧光素酶测定的能力，该荧光素酶测定测试了化合物对早期配子体的作用，并开辟了在配子体发生的整个过程中分析杀配子体化合物的活性的可能性。
The discovery of new antimalarial drugs able to target both the asexual and gametocyte stages of Plasmodium falciparum is critical to the success of the malaria eradication campaign. We have developed and validated a robust, rapid, and cost-effective high-throughput reporter gene assay to identify compounds active against late-stage (stage IV and V) gametocytes. The assay, which is suitable for testing compound activity at incubation times up to 72 h, demonstrates excellent quality and reproducibility, with average Z' values of 0.85 +/- 0.01. We used the assay to screen more than 10,000 compounds from three chemically diverse libraries. The screening outcomes highlighted the opportunity to use collections of compounds with known activity against the asexual stages of the parasites as a starting point for gametocytocidal activity detection in order to maximize the chances of identifying gametocytocidal compounds. This assay extends the capabilities of our previously reported luciferase assay, which tested compounds against early-stage gametocytes, and opens possibilities to profile the activities of gametocytocidal compounds over the entire course of gametocytogenesis.