Flow cytometry to evaluate Theileria sergenti parasitemia using the fluorescent nucleic acid stain, SYTO16.
Flow cytometry to evaluate Theileria sergenti parasitemia using the fluorescent nucleic acid stain, SYTO16.
复制标题
使用荧光核酸染色剂 SYTO16 进行流式细胞术评估塞氏泰勒虫寄生虫血症。
DOI：
--
复制
发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Y. Chikayama
中科院分区：
文献类型：
--
作者：
Yukio Yagi;H. Shiono;Nobuaki Kurabayashi;Kazuhiro Yoshihara;Y. Chikayama
背景
虽然瑟氏泰勒虫的感染是通过这种寄生虫的红细胞内定位来证明的，但为了确定这一点，需要大量的时间和劳动。我们应用流式细胞术检测T。用荧光核酸染色法检测血吸虫病。
方法
从感染T. sergenti用膜渗透性荧光核酸染色剂SYTO 16和氢乙啶（HE）染色。通过配备有在488 nm下操作的单个氩激光器的流式细胞仪测量染色的寄生红细胞。
结果
在FL 1（525 nm）上检测到SYTO 16染色的红细胞内寄生虫，并且寄生的细胞与未寄生的细胞完全分离。然而，HE染色的红细胞不能清楚地分为寄生和未寄生的细胞。SYTO 16染色的寄生虫可重复检测，百分比高于0.1%。通过缩小红细胞部分的光散射门，从分析中消除由CD 18阳性细胞指示的污染性白细胞。SYTO 16阳性细胞百分率与放牧牛寄生虫血症呈显著正相关（r = 0.983）。
结论
SYTO-16流式细胞仪是一种快速、可靠的血吸虫病监测方法。感染了瑟氏病毒的牛
BACKGROUND
Although the infection of Theileria sergenti is demonstrated by intraerythrocytic localization of this parasite, much time and labor are necessary in order to determine this. We applied flow cytometry to evaluate T. sergenti parasitemia using the fluorescent nucleic acid stain method.
METHODS
Peripheral blood samples from cattle infected with T. sergenti were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16, and hydroethidine (HE). Stained parasitized erythrocytes were measured by a flow cytometer equipped with a single argon laser operating at 488 nm.
RESULTS
SYTO16-stained intraerythrocytic parasites were detected on the FL1 (525 nm) and parasitized cells were separated completely from unparasitized cells. However, HE-stained erythrocytes could not be divided clearly into parasitized and unparasitized cells. SYTO16-stained parasites were reproducibly detected at a percentage above 0.1%. Contaminating leukocytes, which were indicated by CD18-positive cells, were eliminated from the analysis by narrowing the light scatter gate of the erythrocyte fraction. A correlation (r = 0.983) between the percentage of SYTO16-positive cells and parasitemia in grazing cattle was observed.
CONCLUSIONS
Flow cytometric detection using SYTO16 is a rapid and reliable method of monitoring parasitemia in T. sergenti-infected cattle.