Plasmodium falciparum: Development and validation of a measure of intraerythrocytic growth using SYBR Green I in a flow cytometer
Plasmodium falciparum: Development and validation of a measure of intraerythrocytic growth using SYBR Green I in a flow cytometer
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DOI：
10.1016/j.exppara.2008.10.008
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发表时间：
2009-02-01
影响因子：
2.1
通讯作者：
Asahi, Hiroko
中科院分区：
文献类型：
--
作者：
Izumiyama, Shinji;Omura, Mako;Asahi, Hiroko
测试促进疟原虫生长和抗疟功效的可靠分析技术非常重要。流式细胞术（FCM）提供了可能性，研究疟疾寄生虫的红细胞内生长的发育阶段，使用核酸染色。为了分析恶性疟原虫的生长，引入SYBR绿色I作为用于氩激光的488 nm线的FCM的嵌入染料。采用FCM的程序，包括固定剂，染料浓度，稀释缓冲液，染色时间，优化，以简化方法。如本文所述的FCM允许根据DNA含量定量寄生虫血症和不同阶段的寄生虫。流式细胞仪和姬姆萨法测定的红细胞寄生率与乳酸脱氢酶法测定的红细胞寄生率一致。该协议扩展到裂殖子计数作为寄生虫生长抑制的敏感测定。(C)2008年爱思唯尔公司All rights reserved.
Reliable analytical techniques to test growth-promoting and antimalarial efficacy on plasmodia are very important. Flow cytometry (FCM) offers the possibility to study developmental stages of intraerythrocytic growth of malaria parasites using nucleic acid staining. To analyze the growth of Plasmodium falciparum SYBR Green I was introduced as an intercalating dye with FCM for the 488 nm line of an argon laser. Procedures employing FCM, including fixatives, dye concentrations, dilution buffer, and staining period, were optimized to simplify the method. FCM as described here allows parasitemia and parasites of different stages to be quantified according to the DNA content. The proportion of parasitized erythrocytes estimated by FCM and the Giemsa method agreed with determination by parasite lactate dehydrogenase. The protocol was extended to merozoite counting as a sensitive assay of growth inhibition of the parasite. (C) 2008 Elsevier Inc. All rights reserved.