Bahamontes-Rosa Noemí, Rodríguez-Alejandre Ane, González-del-Rio Rubén, García-Bustos José Francisco, Mendoza-Losana Alfonso
Diseases of the Developing World, GlaxoSmithKline, 28760 Tres Cantos, Madrid, Spain.
Mol Biochem Parasitol. 2012 Feb;181(2):171-7. doi: 10.1016/j.molbiopara.2011.11.003. Epub 2011 Nov 17.
In order to maximise compliance, the future antimalarial treatment should ideally require just a single-dose administration. This, in turn, demands new fast-acting effective drugs. Currently, methods to measure the in vitro killing rate of antimalarials are based on parasite growth. We have developed and validated a method to determine and classify antimalarial agents based on their cidal or static activity following quantitative Real Time PCR (RT-PCR) analysis. The method described here is a fast, reliable and user-friendly technique with a medium throughput. Metabolic activity of the parasite is followed by measuring mRNA expression levels of several genes during 5 parasite life cycles. mRNA from the parasite culture is then retrotranscribed to cDNA and quantified by RT-PCR. This new method provides a rapid and reproducible way to accurately measure the antimalarial activity of new compounds in vitro against Plasmodium falciparum.
为了最大限度地提高依从性，未来的抗疟治疗理想情况下应只需单剂量给药。这反过来又需要新的速效有效药物。目前，测量抗疟药体外杀灭率的方法是基于寄生虫生长。我们已经开发并验证了一种基于定量实时聚合酶链反应（RT-PCR）分析后的杀疟活性或静态活性来确定和分类抗疟药物的方法。这里描述的方法是一种快速、可靠且用户友好的技术，具有中等通量。在5个寄生虫生命周期中，通过测量几个基因的mRNA表达水平来跟踪寄生虫的代谢活性。然后将来自寄生虫培养物的mRNA逆转录为cDNA，并通过RT-PCR进行定量。这种新方法提供了一种快速且可重复的方式，以准确测量新化合物体外对恶性疟原虫的抗疟活性。
