Development of a highly sensitive, quantitative, and rapid detection system for Plasmodium falciparum-infected red blood cells using a fluorescent blue-ray optical system
Development of a highly sensitive, quantitative, and rapid detection system for Plasmodium falciparum-infected red blood cells using a fluorescent blue-ray optical system
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DOI：
10.1016/j.bios.2019.02.064
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发表时间：
2019-05-01
影响因子：
12.6
通讯作者：
Kataoka, Masatoshi
中科院分区：
文献类型：
--
作者：
Yamamoto, Takeki;Yatsushiro, Shouki;Kataoka, Masatoshi
一个高灵敏度的诊断系统，以确定低密度感染，是由传统的方法错过了检测恶性疟原虫的载体是必要的。研制了一种基于聚碳酸酯扫描盘的荧光蓝光光学系统，用于检测恶性疟原虫感染的红细胞（Pf-iRBC），可同时检测9个样本。使用培养的恶性疟原虫菌株3D 7来检查该系统用于诊断疟疾的潜力。在将RBC悬浮液施加到圆盘上之后，通过旋转将细胞分散在圆盘上。在10分钟的静置期间，使RBC沉降在盘表面上，同时用预先吸附在盘表面上的核荧光染色染料Hoechst 34580对细胞进行染色。通过瞬时旋转去除多余细胞，将RBC在圆盘表面上排列为单层。超过110万个RBC保留在圆盘上用于荧光分析。一个便携式的，电池驱动的荧光图像读取器检测荧光阳性红细胞约40分钟。一个良好的相关性检查Giemsa染色的红细胞通过光学显微镜和开发的系统被证明在寄生虫血症范围为0.0001-1.0%的线性回归分析（R2 = 0.99993）。方法的检出限为0.00020%，重复性好。证明了所开发的系统能够检测亚显微低密度Pf-iRBC并提供准确的定量评价，操作简单。
A highly sensitive diagnostic system for determining low-density infections that are missed by conventional methods is necessary to detect the carriers of Plasmodium falciparum. A fluorescent blue-ray optical system with a polycarbonate scan disc was developed to detect P. falciparum-infected red blood cells (Pf-iRBCs), and nine samples could be analyzed simultaneously. The cultured P. falciparum strain 3D7 was used to examine the potential of the system for diagnosing malaria. After an RBC suspension had been applied to the disc, the cells were dispersed on the disc by rotation. During the 10 min standing period to allow the RBCs to settle on the disc surface, the cells were simultaneously stained with nuclear fluorescence staining dye Hoechst 34580, which was previously adsorbed on the disc surface. RBCs were arranged on the disc surface as a monolayer by removing excess cells through momentary rotation. Over 1.1 million RBCs remained on the disc for fluorescence analysis. A portable, battery-driven fluorescence image reader was employed to detect fluorescence-positive RBCs for approximately 40 min. A good correlation between examination of Giemsa-stained RBCs by light microscopy and the developed system was demonstrated in the parasitemia range of 0.0001-1.0% by linear regression analysis (R2 = 0.99993). The limit of detection of 0.00020% and good reproducibility for parasitemia determination were observed. The ability of the developed system to detect sub-microscopic low-density Pf-iRBCs and provide accurate quantitative evaluation with easy operation was demonstrated.