Establishment of novel in vitro assay for evaluation of killing effect of chloroquine against malaria parasite

根据先前研究(Paguio，Mol Biochem Parasitol，2011)的方法，使用实验室培养的标准菌株氯喹敏感株(HB3)和抗药性寄生虫DD2(东南亚来源)和7G8(南美来源)完成了确认氯喹杀寄生虫效果的第一步后，下一步是使用从乌干达现场回收的冷冻保存的自然寄生虫来验证氯喹的杀寄生虫效果。首先，恢复冷冻保存的乌干达自然寄生虫，并使其适应实验室中的连续培养。一旦成功，就进行氯喹细胞杀灭药物试验。去年，寄生虫的恢复和适应培养失败，因为用过的寄生虫被收集在EDTA中，阻碍了寄生虫的生长。2022年2月至3月和2022年10月至11月，我们在乌干达完成了两次实地调查，收集了135株新鲜寄生虫分离物(2022年2月至3月29株，2022年10月至11月106株)。冷冻保存的寄生虫已从乌干达运往日本。这些野外寄生虫的恢复和适应持续培养的工作正在进行中。

After completing the first step of confirming the chloroquine parasiticidal effect using the laboratory cultured standard strains: chloroquine sensitive (HB3) and resistant parasites: Dd2 (Southeast Asian origin) and 7G8 (South American origin) according to the method of a previous study (Paguio, Mol Biochem Parasitol 2011), the next step was to verify the parasiticidal chloroquine effect using cryopreserved natural parasites recovered from Uganda field site. First by recovering cryopreserved field natural uganda parasites and adapt them to continuous culture in the laboratory. Once successful then perform chloroquine cytocidal drug assay.Last year, parasite recovery and adaptation into culture failed because the used parasites had been collected in EDTA that hindered parasite growth. We changed the protocol to collect fresh parasite isolates in ACD-A that supports parasite growth.From Feburary-March 2022 and October-November 2022, two field surveys have been completed in Uganda and 135 fresh parasite isolates (29 in Feb-March 2022 and 106 in Oct-Nov 2022) have been collected. The cryopreserved parasites have been shipped from Uganda to Japan. Parasite recovery and adaptation of these field parasites into continuous culture is ongoing.