Hashimoto Muneaki, Yokota Kazumichi, Kajimoto Kazuaki, Matsumoto Musashi, Tatsumi Atsuro, Yamamoto Kenichi, Hyodo Tomonori, Matsushita Kiichiro, Minakawa Noboru, Mita Toshihiro, Oka Hiroaki, Kataoka Masatoshi
Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2217-14, Hayashi-cho, Takamatsu, Kagawa 761-0301, Japan.
Konica Minolta, 1 Sakura-mashi, Hino, Tokyo 191-8511, Japan.
Microorganisms. 2020 Sep 4;8(9):1356. doi: 10.3390/microorganisms8091356.
The microscopic examination of Giemsa-stained thin and/or thick blood films (Giemsa microscopy) is the standard method of malaria diagnosis. However, the results of the diagnosis significantly depend on the skills of clinical technicians. Furthermore, sample preparation and analysis are laborious and time-consuming. Therefore, in this study, we investigated if a commercially available fluorescent cell counter, LUNA-FL, was useful for the detection of parasite and the estimation of parasitemia. Whole blood samples from uninfected persons, spiked with -infected erythrocytes, were analysed. Most of the leucocytes and platelets were removed from whole blood samples with SiO-nanofiber filters set on spin columns. The filtered samples were stained with acridine orange, and automatic detection, as well as counting of erythrocytes and parasites, were performed using LUNA-FL. Whole blood, with various levels of parasites, was analysed by Giemsa microscopy or with LUNA-FL to estimate parasitemia, and a comparative analysis was performed. The coefficient determination value of the regression line was high ( = 0.98), indicating that accurate quantitative parasite detection could be performed using LUNA-FL. LUNA-FL has a low running cost; it is compact, fast, and easy to operate, and may therefore be useful for point-of-care testing in the endemic areas.
对吉姆萨染色的薄血膜和/或厚血膜进行显微镜检查(吉姆萨显微镜检查)是疟疾诊断的标准方法。然而,诊断结果在很大程度上取决于临床技术人员的技能。此外,样本制备和分析既费力又耗时。因此,在本研究中,我们调查了一种市售的荧光细胞计数器LUNA-FL是否可用于检测疟原虫和估计疟原虫血症。对添加了感染红细胞的未感染人员的全血样本进行了分析。使用置于旋转柱上的SiO纳米纤维过滤器从全血样本中去除了大部分白细胞和血小板。将过滤后的样本用吖啶橙染色,并使用LUNA-FL进行红细胞和疟原虫的自动检测及计数。对含有不同疟原虫水平的全血进行吉姆萨显微镜检查或使用LUNA-FL分析以估计疟原虫血症,并进行了比较分析。回归线的决定系数值很高(=0.98),表明使用LUNA-FL可以进行准确的疟原虫定量检测。LUNA-FL运行成本低;它体积小巧、速度快且易于操作,因此可能对流行地区的即时检测有用。