流式细胞仪在快速测定鲤春病毒血症病毒滴度中的应用  

Flow Cytometry Assay for Titrating Spring Viraemia of Carp Virus

作　　者：王津津[1] 贾鹏[1] 史秀杰 于力[1] 阮周曦[1] 郑晓聪[1] 何俊强[1] 兰文升[1] 宋思静 刘荭[1] 

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518045

出　　处：《中国动物检疫》2015年第9期82-86,共5页China Animal Health Inspection

基　　金：国家质检总局科技计划项目(2014IK236);国家质检总局公益性行业科研专项(201210055和20120214);深圳检验检疫局科技计划项目(SZ2014202)

摘　　要：本研究建立了流式细胞仪快速检测鲤春病毒血症病毒(spring viraemia of carp virus,SVCV)滴度的方法。运用荧光激活细胞分选(fluorescence-activated cell sorting,FACS)技术检测SVCV A1株对草鱼性腺细胞系(GCO)的感染情况。用SVCV病毒单克隆抗体为一抗,FITC标记的羊抗鼠抗体为二抗,运用FACS来检测感染后不同时间点,以及不同病毒接种量的阳性细胞率。感染第3天时为最佳的病毒滴度测定时间点,测得SVCV的病毒滴度为8.31×105 FIU/m L,最低检测病毒滴度为(1000 FIU/m L),与传统空斑试验(plaque assay,PA)相比,两种方法测得的结果基本一致。实验结果表明,FACS是一种简捷、高效、直接的检测SVCV滴度的方法,是一种新型的病毒滴度测定方法。A flow cytometry assay was established for rapid titrating spring viraemia of carp virus （SVCV） . Fluores- cence-activated cell sorting （FACS） was used to detect SVCVA1 strain-infected GCO cells. The infected cells reacted with anti-SVC monoclonal antibody before being covered with labeled fluorescein-anti-mouse immunoglobulin. FACS was used to analyze the percentage of infected cells at different time points and different virus infections. The results demonstrate that day 3 was the best time to titer SVCV. The titers of SVCV A 1 strain was 8.31× 10 ^5 FIU/mL. The lowest limit of detection by FACS was 1000 infectious units per milliliter of inoculum. Comparing the titers of SVCV A1 strain with FACS and plaque assay, similar titers were obtained with the two assays. The novel FACS is recommended as a simple, efficient and direct assay for titrating SVCV and other virus.

关 键 词：鲤春病毒血症 草鱼性腺细胞系 流式细胞术 空斑试验 病毒滴度 

分 类 号：S941.416[农业科学—水产养殖]