中国海洋大学学报（自然科学版）2012，Vol.42Issue(5)：55-59,5.
免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度
Development of a Virus Titration Assay for Lymphocystis Disease Virus by an Immunocytochemical Staining Method
摘要
Abstract
Abstract: An immunocytochemical staining method was developed for virus titration assay of lymphocys-tis disease virus (LCDV) in cultures of a flounder gill (FG) cell line. FG cells were propagated into a 48 well tissue culture plate and post inoculated with two-fold serially diluted LCDV, purified from lympho-cystis cells of Japanese flounder Paralichthys olivaceus. After infection, the cell cultures were fixed and incubated with LCDV-specific monoclonal antibody in combination with biotin-avidin system. Infected cells and uninfected cells, stained with alkaline phosphatase substrate, could be differentiated by presence and absence of red color in the cytoplasm. Each positive well was scored microscopically based on the appearance of immunostained cells and the titer of the LCDV infectivity in FG cells was measured to be 1. 77×210TCID50 per mL by the immunocytochemical staining method. The immunocytochemical staining method is proved to be accurate and sensitive for measuring the titer of LCDV.关键词
淋巴囊肿病毒/牙鲆鳃细胞/组织细胞培养半数感染量/免疫细胞化学法Key words
lymphocystis disease virus/ flounder gill cells/ TCID50/ immunocytochemical staining method分类
医药卫生引用本文复制引用
宋佳蕾,绳秀珍,战文斌..免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度[J].中国海洋大学学报（自然科学版）,2012,42(5):55-59,5.基金项目
国家自然科学基金项目(31172429 （）
31072232) （）
国家高技术研究发展计划项目(2006AA100306)资助 （2006AA100306）
