流式细胞术检测对虾白斑症病毒(WSSV)对克氏原螯虾血细胞的感染  

Detection of WSSV in Circular Haemocytes of Crayfish by Flow Cytometry

作　　者：王轶南[1] 战文斌[1] 邢婧[1] 

机构地区：[1]中国海洋大学教育部海水养殖重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2008年第3期384-388,共5页Periodical of Ocean University of China

基　　金：国家重点基础研究发展计划(2006CB101806);国家高技术研究发展计划项目(2006AA100312)资助

摘　　要：用WSSV粗提液人工感染克氏原螯虾,感染后分别在6,12,18,24h采血分离血细胞,通过WSSV的混合单抗和FITC标记的羊抗鼠二抗的间接免疫荧光染色,应用流式细胞仪检测病毒对克氏原螯虾血细胞的感染并分析感染强度。结果表明:分离获得的血细胞形态完好,感染后血细胞内的病毒被标记上了绿色荧光;流式检测发现,随感染时间延长,被感染细胞的百分率及平均荧光强度均逐渐升高,反映血细胞的病毒感染程度逐渐加深;PCR检测同时证实人工感染后各采样点血细胞样品均为WSSV阳性。Crayfish（Cambarus proclarkii） were artificially infected with WSSV solution and the haemocytes were collected at 6 h, 12 h, 18 h and 24 h post-infection and stained with WSSV specific Mabs and secondary antibody conjugated with FITC, a flow cytometer was used to detect WSSV and analyse the infection degree of crayfish haemocytes. The results showed that the collected cells were intact in shape, the virus within the cells were stained with green fluorescence and could be detected by FCM. Furthermore, the percent and mean fluorescence intensity of infected cells increased from 6 h to 24 h post-infection, which indicated the deepening of virus infection in crayfish haemocytes. PCR confirmed the haemocytes samples after virus infection were WSSV positive.

关 键 词：克氏原螯虾 血细胞 对虾白斑症病毒(WSSV) 流式细胞术 

分 类 号：S945.19[农业科学—水产养殖]