鳜鱼病毒病原及细胞感染特性的研究
作者：方勤 艾桃山 邹桂平 喻运珍 刘家欣 汪亚平
单位：方勤(中国科学院水生生物研究所，淡水生态与生物技术国家重点实验室，武汉 430072；中科学院武汉病毒研究所，武汉 430071)；艾桃山 喻运珍(武汉市水产科学研究所，武汉 430006)；邹桂平 刘家欣(中国科学院武汉病毒研究所，武汉 430071)；汪亚平(中国科学院水生生物研究所，淡水生态与生物技术国家重点实验室，武汉 430072)
关键词：鳜鱼；病毒；敏感细胞系
中国病毒学000315摘要：在患暴发流行性发病鳜鱼的脾、肾、肝、鳃组织细胞的超薄切片中观察到直径为120-150 nm的二十面体病毒粒子，其截面呈六角形，有囊膜。将病鱼内脏与10倍体积的PBS缓冲液制成组织匀浆液，过滤除菌后感染对数生长期的草鱼肾细胞。结果表明，该感染液在28 ℃条件下，培养36-48 h，生长旺盛的草鱼肾细胞开始出现CPE，5-7 d感染的单层细胞逐渐脱落死亡。该细胞病毒悬液具有感染性，可持续稳定地进行传代培养。感染细胞的病毒培养液，经差异离心纯化，负染制片，电镜下观察到大量的病毒粒子，对照培养物中未发现任何病毒颗粒。
, http://www.100md.com
中图分类号：Q939.4， S965.117 文献标识码：A
文章编号：1003-5125(2000)03-0297-05
Study of the Siniperca chuatsi Viral Pathogen and its
Infectious Characteristics in Fish Cells
FANG Qin
(State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology,Chinese Academy of Sciences, Wuhan 430072, China；
, 百拇医药
Wuhan Institute of Virology, Academia Sinica, Wuhan 430071, China)
WANG Ya-ping
(State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology,Chinese Academy of Sciences, Wuhan 430072, China)
ZOU Gui-ping，LIU Jia-xin
(Wuhan Institute of Virology, Academia Sinica, Wuhan 430071, China)
AI Tao-shan，YU Yun-zhen
, http://www.100md.com
(Wuhan Institute of fishary, Wuhan 430006, China)
Abstract: An icosahedral virus was found in the ultra-thin section of spleen, kidney, liver, gill of the outbreak-infectious Siniperca chuatsi by electron microscope. The virions of section show hexagon, about 125-150 nm in diameter, and have envelope outside. Cell culture indicated that CIK cell was sensitive to the virus, which was isolated from the internal organs of Siniperca chuatsi. The CPE can be seen after being infected for 36-48 hours at 28℃. Cell cultured virus suspension is infectious and can be subcultured steadily in CIK cell line. Many virions isolated by different centrifugation from infected cell culture can be observed by negative stained electron microscope method, but no virions were detected in the control samples.℃. Cell cultured virus suspension is infectious and can be subcultured steadily in CIK cell line. Many virions isolated by different centrifugation from infected cell culture can be observed by negative stained electron microscope method, but no virions were detected in the control samples.
, 百拇医药
Key words: Siniperca chuatsi; Virus; Sensitive cell line
基金项目：“淡水生态与生物技术国家重点实验室”开放项目(2000FB07)；湖北省自然科学
基金资助项目(99J097)。
作者简介：方勤(1961年-)，女，湖北省人，副研究员。主要从事水生动物病毒的分子生物学及基因工程研究。
参考文献
[1] 陈昌福，史维舟，赵桂珍，等.翘嘴鳜烂腮病病原菌的分离及初步鉴定[J].华中农业大学学报，1995，14(2)：263-266
[2] 吴淑勤，李新辉，潘厚军，等.鳜暴发性传染病病原研究[J].水产学报，1997，21(增刊)：56-60
, 百拇医药
[3] 何建国，翁少萍，黄志坚，等.鳜暴发流行病病毒性病原的研究[J].中山大学学报(自然科学版)，1998，37(5)：77-77
[4] 李新辉，吴淑勤，潘厚军，等.一种检测鳜鱼病毒方法[J]，中国水产科学.1997，4(5)：112-114
[5] 方勤，柯丽华，蔡宜权，等.草鱼出血病病毒的生长特性及高滴价培养[J].病毒学杂志，1989，3：315-319
[6] 杨复华. 病毒学[M].长沙：湖南科学技术出版社，1992.453-482
[7] Rodger H D, Knobs M, Macartney A, et al. Systemic iridovirus infection in freshwater angelifish, Pterophyllum scalare (Lichtenstein) [J]. Journal of Fish Disease, 1997,20:69-70
, 百拇医药
[8] Langdon J S, Humphrey J D, Williams L M. First virus isolation from Australian fish: an iridovirus-like virus pathogen from redfin perch. Perca fluviatilis L [J]. Journal of Fish Disease, 1986,9:263-268
[9] McGrogan D G, Ostland V E, Byrne P J, et al. Systemic disease involving an iridovirus-like agent in cultured tilapia. Oreochromis niloticus L. -a case report [J]. Journal of Fish Diseases, 1998,21:149-152
[10] Langdon J S, Humphrey J D, Williams L M. Outbreaks of an EHNV-like iridovirus in cultured rainbow trout, Salmo gaidneri Richardson, in Australia [J]. Journal of Fish Diseases, 1988,11:92-96
[11] Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning, A Laboratory Manual [M]. 2nd ed. USA, NY: Cold Spring Harbor, 1989. 954-956
收稿日期：1999-06-28，修回日期：2000-01-13, 百拇医药
单位：方勤(中国科学院水生生物研究所，淡水生态与生物技术国家重点实验室，武汉 430072；中科学院武汉病毒研究所，武汉 430071)；艾桃山 喻运珍(武汉市水产科学研究所，武汉 430006)；邹桂平 刘家欣(中国科学院武汉病毒研究所，武汉 430071)；汪亚平(中国科学院水生生物研究所，淡水生态与生物技术国家重点实验室，武汉 430072)
关键词：鳜鱼；病毒；敏感细胞系
中国病毒学000315摘要：在患暴发流行性发病鳜鱼的脾、肾、肝、鳃组织细胞的超薄切片中观察到直径为120-150 nm的二十面体病毒粒子，其截面呈六角形，有囊膜。将病鱼内脏与10倍体积的PBS缓冲液制成组织匀浆液，过滤除菌后感染对数生长期的草鱼肾细胞。结果表明，该感染液在28 ℃条件下，培养36-48 h，生长旺盛的草鱼肾细胞开始出现CPE，5-7 d感染的单层细胞逐渐脱落死亡。该细胞病毒悬液具有感染性，可持续稳定地进行传代培养。感染细胞的病毒培养液，经差异离心纯化，负染制片，电镜下观察到大量的病毒粒子，对照培养物中未发现任何病毒颗粒。
, http://www.100md.com
中图分类号：Q939.4， S965.117 文献标识码：A
文章编号：1003-5125(2000)03-0297-05
Study of the Siniperca chuatsi Viral Pathogen and its
Infectious Characteristics in Fish Cells
FANG Qin
(State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology,Chinese Academy of Sciences, Wuhan 430072, China；
, 百拇医药
Wuhan Institute of Virology, Academia Sinica, Wuhan 430071, China)
WANG Ya-ping
(State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology,Chinese Academy of Sciences, Wuhan 430072, China)
ZOU Gui-ping，LIU Jia-xin
(Wuhan Institute of Virology, Academia Sinica, Wuhan 430071, China)
AI Tao-shan，YU Yun-zhen
, http://www.100md.com
(Wuhan Institute of fishary, Wuhan 430006, China)
Abstract: An icosahedral virus was found in the ultra-thin section of spleen, kidney, liver, gill of the outbreak-infectious Siniperca chuatsi by electron microscope. The virions of section show hexagon, about 125-150 nm in diameter, and have envelope outside. Cell culture indicated that CIK cell was sensitive to the virus, which was isolated from the internal organs of Siniperca chuatsi. The CPE can be seen after being infected for 36-48 hours at 28℃. Cell cultured virus suspension is infectious and can be subcultured steadily in CIK cell line. Many virions isolated by different centrifugation from infected cell culture can be observed by negative stained electron microscope method, but no virions were detected in the control samples.℃. Cell cultured virus suspension is infectious and can be subcultured steadily in CIK cell line. Many virions isolated by different centrifugation from infected cell culture can be observed by negative stained electron microscope method, but no virions were detected in the control samples.
, 百拇医药
Key words: Siniperca chuatsi; Virus; Sensitive cell line
基金项目：“淡水生态与生物技术国家重点实验室”开放项目(2000FB07)；湖北省自然科学
基金资助项目(99J097)。
作者简介：方勤(1961年-)，女，湖北省人，副研究员。主要从事水生动物病毒的分子生物学及基因工程研究。
参考文献
[1] 陈昌福，史维舟，赵桂珍，等.翘嘴鳜烂腮病病原菌的分离及初步鉴定[J].华中农业大学学报，1995，14(2)：263-266
[2] 吴淑勤，李新辉，潘厚军，等.鳜暴发性传染病病原研究[J].水产学报，1997，21(增刊)：56-60
, 百拇医药
[3] 何建国，翁少萍，黄志坚，等.鳜暴发流行病病毒性病原的研究[J].中山大学学报(自然科学版)，1998，37(5)：77-77
[4] 李新辉，吴淑勤，潘厚军，等.一种检测鳜鱼病毒方法[J]，中国水产科学.1997，4(5)：112-114
[5] 方勤，柯丽华，蔡宜权，等.草鱼出血病病毒的生长特性及高滴价培养[J].病毒学杂志，1989，3：315-319
[6] 杨复华. 病毒学[M].长沙：湖南科学技术出版社，1992.453-482
[7] Rodger H D, Knobs M, Macartney A, et al. Systemic iridovirus infection in freshwater angelifish, Pterophyllum scalare (Lichtenstein) [J]. Journal of Fish Disease, 1997,20:69-70
, 百拇医药
[8] Langdon J S, Humphrey J D, Williams L M. First virus isolation from Australian fish: an iridovirus-like virus pathogen from redfin perch. Perca fluviatilis L [J]. Journal of Fish Disease, 1986,9:263-268
[9] McGrogan D G, Ostland V E, Byrne P J, et al. Systemic disease involving an iridovirus-like agent in cultured tilapia. Oreochromis niloticus L. -a case report [J]. Journal of Fish Diseases, 1998,21:149-152
[10] Langdon J S, Humphrey J D, Williams L M. Outbreaks of an EHNV-like iridovirus in cultured rainbow trout, Salmo gaidneri Richardson, in Australia [J]. Journal of Fish Diseases, 1988,11:92-96
[11] Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning, A Laboratory Manual [M]. 2nd ed. USA, NY: Cold Spring Harbor, 1989. 954-956
收稿日期：1999-06-28，修回日期：2000-01-13, 百拇医药