Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus

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	Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus
	Zhou, Guang-Zhou; Li, Zheng-Qiu; Zhang, Qi-Ya
刊名	VIRAL IMMUNOLOGY
	2006-12-01
卷号	19 期号:4 页码:637-645
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
ISSN号	0882-8245
通讯作者	Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
中文摘要	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.
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.
学科主题	Immunology; Virology
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Immunology ; Virology
研究领域[WOS]	Immunology ; Virology
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
收录类别	SCI
语种	英语
WOS记录号	WOS:000243346600006
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/8730]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
作者单位	Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
推荐引用方式 GB/T 7714	Zhou, Guang-Zhou,Li, Zheng-Qiu,Zhang, Qi-Ya. Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus[J]. VIRAL IMMUNOLOGY,2006,19(4):637-645.
APA	Zhou, Guang-Zhou,Li, Zheng-Qiu,&Zhang, Qi-Ya.(2006).Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus.VIRAL IMMUNOLOGY,19(4),637-645.
MLA	Zhou, Guang-Zhou,et al."Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus".VIRAL IMMUNOLOGY 19.4(2006):637-645.