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Development of new malaria parasite detection system : Establishment of automatic and simple system

新型疟疾寄生虫检测系统的开发：建立自动化且简易的系统

基本信息

批准号：
10557030
负责人：
SAITO Atsuko
金额：
$ 3.01万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557030/
关键词：
malaria Plasmodium flow cytometry Diagnosis acridine orange hemolysis 蛍光染色

项目摘要

We established a simple and sensitive flow cytometric detection system for Plasmodium falciparum that involves lysing erythrocytes and staining parasites at the same time using a newly developed hemolysing solution containing acridine orange. In this system, freed parasites of P.falciparum could be plotted separately from erythrocyte ghosts, white blood cells and platelets on the two-dimensional scattergram of forward angle light scatter and green fluorescence by flow cytometry with an argon laser. It took only 2-3 minutes per sample to obtain the scattergram and analyze the data, including the time of sample preparation for the flow cytometric analysis. Sample preparation does not require any particularly difficult handling procedures. The threshold of parasite detection was almost equal to that of microscopic examination for cultured P.falciparum. The results of drug susceptibility assays using this system were quite similar to those obtained using microscopic examination. In this system, parasites at different erythrocytic stages could be easily distinguished. Moreover, using a flow cytometer with imaging device, clear images of gametocytes were obtained. This system has not only the potential to be useful for laboratory studies, but also a high potential of becoming an ideal new diagnostic system for malaria in the near future.

采用新研制的吖啶橙溶血液，建立了一种简便、灵敏的恶性疟原虫红细胞溶血和寄生虫染色的流式细胞术检测系统。在该系统中，利用氩激光流式细胞术可以在正角光散射和绿色荧光的二维散点图上将释放的恶性疟原虫与红细胞鬼影、白细胞和血小板分开绘制。每个样品获得散点图和分析数据只需要2-3分钟，包括流式细胞术分析的样品制备时间。样品制备不需要任何特别困难的处理程序。培养恶性疟原虫的检出阈值与镜检阈值基本相等。该系统的药敏试验结果与镜检结果基本一致。在这个系统中，很容易区分不同红细胞阶段的寄生虫。利用带显像装置的流式细胞仪，获得了配子体清晰的图像。该系统不仅有潜力用于实验室研究，而且在不久的将来也很有可能成为一种理想的新的疟疾诊断系统。