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学科主题: Immunology; Virology
题名: Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus
作者: Zhou, Guang-Zhou; Li, Zheng-Qiu; Zhang, Qi-Ya
通讯作者: Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
关键词: HEMATOPOIETIC NECROSIS VIRUS ; HEMORRHAGIC SEPTICEMIA VIRUS ; COMPLETE GENOMIC SEQUENCE ; SCALLOP CHLAMYS-FARRERI ; FLOW-CYTOMETRY ; PARALICHTHYS-OLIVACEUS ; FISH RHABDOVIRUS ; SPRING VIREMIA ; DISEASE-VIRUS ; MARINE FISH
刊名: VIRAL IMMUNOLOGY
发表日期: 2006-12-01
DOI: 10.1089/vim.2006.0049
卷: 19, 期:4, 页:637-645
收录类别: SCI
文章类型: Article
部门归属: Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Immunology ; Virology
研究领域[WOS]: Immunology ; Virology
摘要: Five monoclonal antibodies (mAbs), 1G8, 1H9, 2D2, 2D3, and 2F5, against Scophthalmus maximus rhabdovirus (SMRV) were prepared. Characterization of the mAbs included indirect enzyme-linked immunosorbent assay, isotyping, viral inhibition assay, immunofluorescence staining of virus-infected cell cultures, and Western blot analysis. Isotyping revealed that 1G8 and 1H9 were of the IgG2b subclass and that the other three were IgM. 2D2, 2D3, and 2F5 partially inhibited SMRV infection in epithelioma. papulosum cyprinid (EPC) cell culture. Western blotting showed that all five mAbs could react with two SMRV proteins with molecular masses of approximately 30 kDa (P) and 26 kDa (M). These two proteins were localized within the cytoplasm of SMRV-infected EPC cells by immunofluorescence assay. Also, progressive foci of viral replication in cell cultures were monitored from 6 to 24 h, using mAb 2D3 as the primary antibody. A flow cytometry procedure was used to detect and quantify SMRV-infected (0.01 PFU/cell) EPC cells with mAb 2D3, and 10.8% of cells could be distinguished as infected 36 h postinfection. Moreover, mAb 2D3 was successfully applied for the detection of viral antigen in cryosections from flounder tissues by immunohistochemistry tests.
英文摘要: Five monoclonal antibodies (mAbs), 1G8, 1H9, 2D2, 2D3, and 2F5, against Scophthalmus maximus rhabdovirus (SMRV) were prepared. Characterization of the mAbs included indirect enzyme-linked immunosorbent assay, isotyping, viral inhibition assay, immunofluorescence staining of virus-infected cell cultures, and Western blot analysis. Isotyping revealed that 1G8 and 1H9 were of the IgG2b subclass and that the other three were IgM. 2D2, 2D3, and 2F5 partially inhibited SMRV infection in epithelioma. papulosum cyprinid (EPC) cell culture. Western blotting showed that all five mAbs could react with two SMRV proteins with molecular masses of approximately 30 kDa (P) and 26 kDa (M). These two proteins were localized within the cytoplasm of SMRV-infected EPC cells by immunofluorescence assay. Also, progressive foci of viral replication in cell cultures were monitored from 6 to 24 h, using mAb 2D3 as the primary antibody. A flow cytometry procedure was used to detect and quantify SMRV-infected (0.01 PFU/cell) EPC cells with mAb 2D3, and 10.8% of cells could be distinguished as infected 36 h postinfection. Moreover, mAb 2D3 was successfully applied for the detection of viral antigen in cryosections from flounder tissues by immunohistochemistry tests.
关键词[WOS]: HEMATOPOIETIC NECROSIS VIRUS ; HEMORRHAGIC SEPTICEMIA VIRUS ; COMPLETE GENOMIC SEQUENCE ; SCALLOP CHLAMYS-FARRERI ; FLOW-CYTOMETRY ; PARALICHTHYS-OLIVACEUS ; FISH RHABDOVIRUS ; SPRING VIREMIA ; DISEASE-VIRUS ; MARINE FISH
语种: 英语
WOS记录号: WOS:000243346600006
ISSN号: 0882-8245
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/8730
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China

Recommended Citation:
Zhou, Guang-Zhou; Li, Zheng-Qiu; Zhang, Qi-Ya.Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus,VIRAL IMMUNOLOGY,2006,19(4):637-645
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