卡耶坦环孢子虫流式细胞仪检测方法的建立  

Establishment of detection method for Cyclospora cayetanensis by flow cytometry

作　　者：李俊强[1,3] 孙芳芳[1,3] 郅玉宝[2] 齐萌[1,3] 武岳[1,3] 罗楠楠[1,3] 黄建营 王荣军[1,3] 宁长申[1,3] 张龙现[1,3] 

机构地区：[1]河南农业大学,牧医工程学院,郑州450002 [2]河南省农业科学院,河南省动物免疫学重点实验室,郑州450002 [3]河南农业大学.河南省人兽共患病国际联合实验室,郑州345000

出　　处：《国际医学寄生虫病杂志》2014年第1期16-19,共4页International JOurnal of Medical Parasitic Diseases

基　　金：基金项目：河南省高校人兽共患病病原生物学科研创新团队（012IRTSTHN005）;国家重大科技专项课题（2012ZX10004-220、2008ZX10004-011）

摘　　要：目的建立流式细胞仪检测卡耶坦环孢子虫（Cyclospora cayetanensis）卵囊的方法。方法将环孢子虫卵囊液和阴性对照液按一定比例混合，分别制备密度为131080．0、26216．0、5243．2、1048．6、209．7、41．9和8．4个，ml的卵囊液，同时设阴性对照组和空白对照组，分别经流式细胞仪筛选检测，每次分析的常量设定为20000个事件．选定聚集区域R1且在488nm光谱下能自发蓝色荧光区域R2的事件进行分析。结果环孢子虫卵囊液密度低于1048．6个／ml时．卵囊事件量与阴性卵囊液中杂质事件量无差异．流式细胞仪对环孢子虫卵囊量的平均检测限为34．0～77．5个。结论流式细胞仪在临床检测患者粪便时．事件发生数在78个以上即可诊断为环孢子虫病。Objective To develop a detection method for Cyclospora cayetanensis by flow cytometry. Methods The density of Cyclospora oocysts was set as 131 080.0, 26 216.0, 5 243.2, 1 048.6, 209.7, 41.9 and 8.4 oocysts/ml, respectively, while the negative control（Cyclospora-negative liquid） and blank control were used. A total of 20 000 events were analyzed each time under the condition of that a cluster（R1 ） representing Cyclospora oocysts could be clearly observed with blue autofluorescence at 488 nm （R2）. Results There were no difference between test group and negative control when the concentration was below 1 048.6 oocysts/ml, therefore, the mean detection limit of the Cyclospora cayetanensis oocysts was between 34.0 to 77.5 by flow cytometry analysis. Conclusion When detecting Cyclospora oocysts in fecal specimens from clinical patients by flow cytometry, samples having more than 78 events were diagnosed as cyclosporasis.

关 键 词：卡耶坦环孢子虫 流式细胞仪 检测 

分 类 号：R531.5[医药卫生—内科学]