Application of flow cytometry to studies of pathogenic free-living amoebae
Application of flow cytometry to studies of pathogenic free-living amoebae
复制标题
流式细胞术在致病性自由生活阿米巴研究中的应用
DOI：
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发表时间：
1982
影响因子：
4.4
通讯作者：
C. Fliermans
中科院分区：
文献类型：
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作者：
L. Muldrow;R. Tyndall;C. Fliermans
奈格里属和阿米巴属的小型、自由生活的变形虫可引起致命的变形虫性脑膜脑炎。以前的研究表明，致病性阿米巴与热改变的水有关。利用免疫荧光和荧光素结合的伴刀豆球蛋白A，已经开发出了从这种水中鉴定致病性和非致病性阿米巴属种的流式细胞术。流式细胞术是用细胞荧光照相机完成的，其中细胞分散在悬浮的载液中，并在聚焦的氩离子激光束前通过。然后通过散射光（大小）或荧光的程度来区分细胞。流式细胞术技术已被证明是有效的环境样品，如鉴定致病性福氏耐格里和非致病性格鲁贝耐格里和卡氏阿米巴从萨凡纳河厂在南卡罗来纳州。细胞荧光分析的环境样品有几个优势，目前的方法分离和分类的自由生活阿米巴。有了这个系统，它是可能的，以快速识别物种和定量混合物的致病性阿米巴在环境样品中。阿米巴分离株的细胞荧光分析减少了目前筛选环境中致病性阿米巴所需的时间。细胞荧光图允许检测和种属鉴定的非嗜热耐格里属。和棘阿米巴属。不能通过常规方法容易地分离用于物种鉴定。与荧光显微镜相比，流式细胞术的其他优点包括由于测量的数量大而具有高度的统计精度、高免疫荧光滴度以及消除主观性和荧光褪色。
Species of small, free-living amoebae of the genera Naegleria and Acanthamoeba can cause fatal amoebic meningoencephalitis. Previous investigations have shown that pathogenic amoebae are associated with thermally altered water. Flow cytometric techniques for identifying species of pathogenic and nonpathogenic amoebae from such water have been developed, using immunofluorescence and fluorescein-bound concanavalin A. Flow cytometry is accomplished with a cytofluorograph, in which cells are dispersed in a suspended carrier liquid and passed in front of a focused argon ion laser beam. Cells are then distinguished by the degree of scattered light (size) or fluorescence. Flow cytometry techniques have proven efficient for environmental samples, as indicated by the identification of pathogenic Naegleria fowleri and nonpathogenic Naegleri gruberi and Acanthamoeba castellanii isolated from the Savannah River Plant in South Carolina. Cytofluorographic analysis of environmental samples has several advantages over the current methods of isolation and classification of free-living amoebae. With this system, it is possible to rapidly identify species and quantitate mixtures of pathogenic amoebae in environmental samples. Cytofluorographic analysis of amoebic isolates reduces the time presently required to screen environmental sites for pathogenic amoebae. The cytofluorograph permits detection and species identification of nonthermophilic Naegleria spp. and Acanthamoeba spp. that could not easily be isolated for species identification by conventional methods. Other advantages of flow cytometry over fluorescent microscopy include a high degree of statistical precision due to the large numbers measured, high immunofluorescent titers, and elimination of subjectivity and fluorescence fading.