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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (18): 457-460.

所属专题： 生物技术； 水产渔业

• 水产 渔业 • 上一篇    下一篇

鱼类淋巴囊肿病毒感染牙鲆鳃细胞系的研究

程顺峰   

  • 收稿日期:2009-07-17 修回日期:2009-07-21 出版日期:2009-09-20 发布日期:2009-09-20
  • 基金资助:

    山东省自然科学基金项目

Study of Lymphocystis Disease Virus Infected to Flounder Gill Cell Line

  • Received:2009-07-17 Revised:2009-07-21 Online:2009-09-20 Published:2009-09-20

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摘要：

摘要：以患病牙鲆的囊肿组织为实验材料，冰裕匀浆后采用低速离心的方法提取淋巴囊肿病毒，用粗提的病毒液感染牙鲆鳃细胞系FG-9307，同时应用以抗淋巴囊肿病毒单克隆抗体为第一抗体的间接免疫荧光抗体技术检测淋巴囊肿病毒是否感染了牙鲆鳃细胞并在其中增殖。结果发现生长旺盛的单层牙鲆鳃细胞接种淋巴囊肿病毒2天后出现明显的细胞病变效应，形成空斑，应用间接免疫荧光抗体方法检测结果为抗淋巴囊肿病毒单抗与未接种淋巴囊肿病毒的牙鲆鳃细胞呈阴性反应，而接种病毒的牙鲆鳃细胞在接种后，细胞质中有明显的特异性荧光，该结果说明牙鲆鳃细胞系是淋巴囊肿病毒的敏感细胞系。

关键词: 萱草, 萱草, 结缕草, 抗旱性, 生理生化指标

Abstract:

Abstract: Lymphocystis disease virus (LCDV), the causative agent of the lymphocystis disease, has resulted in a great economic loss in marine culture industry and become factor restricting aquaculture development. In this article, Flounder gill cell line (FG-9307) was used to confirm susceptive to LCDV. LCDV viral suspension was separated from lymphocystis cells of diseased flounder Paralichthys olivaceus by cell disruption, centrifugation. After passed through 0.45μm filter, the virus suspension was inoculated with FG-9307 cells cultured in 96-well plates. 1-2 days later, FG cells exhibit cytopathic effect (CPE). Indirect immunofluorescence assay test (IFAT) was employed to detect viral infected by using monoclonal antibodies against LCDV. The results of IFAT showed that LCDV was located at the cell membrance immediately after post infection, and then it reached strongest positive in cytosol after post inoculation 48-72 hours. These date proved that FG-9307 cell line was susceptive to LCDV.