Lysercell M enhances the detection of stage-specific Plasmodium-infected red blood cells in the automated hematology analyzer XN-31 prototype
Lysercell M enhances the detection of stage-specific Plasmodium-infected red blood cells in the automated hematology analyzer XN-31 prototype
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DOI：
10.1016/j.parint.2020.102206
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发表时间：
2021-02-01
影响因子：
1.9
通讯作者：
Uchihashi, Kinya
中科院分区：
文献类型：
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作者：
Toya, Yuji;Tougan, Takahiro;Uchihashi, Kinya
全自动血液分析仪XN-30（用于研究）和XN-31原型（用于诊断支持）可以轻松快速地检测疟原虫感染的红细胞（iRBC），并在大约1分钟内区分寄生虫的发育阶段。两种专用试剂Lysercell M和Fluorocell M可与分析仪一起使用。Lysercell M在增强Fluorocell M中核酸染色染料的荧光强度和改变细胞形态方面起着不可或缺的作用。Lysercell M的这些作用已经根据经验确定，但没有充分分析。在这项研究中，Lysercell M的性质进行了分析，使用两个流式细胞仪和荧光显微镜。首先，评价用Lysercell M或磷酸盐缓冲盐水（PBS）处理的iRBC发射的荧光强度。其次，测量用Lysercell M或PBS处理的RBC的大小。最后，使用成像流式细胞仪重建XN-31原型系统的M散点图（细胞图）后观察单个寄生虫的形态。这些分析表明，用Lysercell M处理iRBC可使染色寄生虫核酸的荧光强度增加约10倍，并以阶段特异性方式减小iRBC的大小，从而有助于鉴定和定量环状、滋养体和鞭毛体阶段的iRBC。这些特性表明Lysercell M可用于快速检测iRBC并准确区分寄生虫发育阶段，从而有助于XN-30和XN-31原型分析仪的可用性。
The automated hematology analyzers XN-30 (for research) and XN-31 prototype (for diagnosis support) can easily and rapidly detect Plasmodium-infected red blood cells (iRBCs) and distinguish the developmental stages of the parasite in approximately 1 min. Two dedicated reagents, Lysercell M and Fluorocell M, are available with the analyzers. Lysercell M plays an indispensable role in enhancing the fluorescence intensity of the nucleic acid staining dye in Fluorocell M and altering cell morphology. These effects of Lysercell M have been empirically determined but insufficiently analyzed. In this study, the properties of Lysercell M were analyzed using two flow cytometers and a fluorescence microscope. First, the fluorescence intensity emitted by iRBCs treated with Lysercell M or phosphate-buffered saline (PBS) was evaluated. Second, the size of RBCs treated with Lysercell M or PBS was measured. Finally, the morphology of individual parasites was observed after reconstruction of an M scattergram, a cytogram of the XN-31 prototype system, using an imaging flow cytometer. These analyses showed that treatment of iRBCs with Lysercell M increased the fluorescence intensity of stained parasite nucleic acids by approximately 10-fold and reduced the size of iRBCs in a stage-specific manner, facilitating the identification and quantification of ring form, trophozoite, and schizont stage iRBCs. These properties suggest that Lysercell M is useful for rapidly detecting iRBCs and accurately distinguishing the parasite developmental stages, thereby contributing to the usability of the XN-30 and XN-31 prototype analyzers.