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Development and characterization of monoclonal antibodies to spring viraemia of carp virus
Chen, Zhong-Yuan1; Liu, Hong2; Li, Zheng-Qiu1; Zhang, Qi-Ya1; Zhang, QY, Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
2008-06-15
Source PublicationVETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
ISSN0165-2427
Volume123Issue:3-4Pages:266-276
AbstractFive monoclonal antibodies (mAbs) against spring viraemia of carp (SVCV0504, isolated from common carp in China) were produced from mice immunized with purified virus preparations. The virion of SVCV contains five structural proteins, representing the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (Q. Western blotting analysis revealed that three mAbs (1145, IE10, and 11-17) recognized specifically to a single protein of 47 kDa (N), the mAb 3G4 reacted with, two SVCV0504 proteins of 69 kDa (G) and 47 kDa (N), while the mAb 1A9 reacted with three SVCV0504 proteins of 69 kDa (G), 50 kDa (P), and 47 kDa (N). By indirect ELISA, two mAbs (1H5 and 11-17) showed cross-reactivity with pike fry rhabdovirus (PFRV), but no cross-reactions with the Siniperca chuatsi rhabdovirus (SCRV), Scophthalmus maximus rhabdovirus (SMRV), Paralichthys olivaceus rhabdovirus (PoRV) were demonstrated with the five mAbs. Indirect immunofluorescence showed intense fluorescence in the cytoplasm of the SVCV0504-infected epithelioma papulosum cyprini (EPC) cells in areas corresponding to the location of granular structures. The sucrose gradient-purified SVCV0504 particles could be detected successfully by these mAbs using immunodot blotting. mAb 1A9 could completely neutralize 100 TCID50 (50% tissue culture infective dose) of SVCV0504 at a dilution of 1:8. This is the first report of development of the neutralizing mAbs against SVCV. The mAb 1A9 was analyzed further and could be used to successfully detect viral antigens in the infected-EPC cell cultures or in cryosections from experimentally infected crucian carp (Carassius auratus) by immunohistochemistry assay. Furthermore, a flow cytometry procedure for the detection and quantification of cytoplasmic SVCV0504 in cell cultures was developed with mAb 1A9. At 28 h after inoculation with the virus (0.01 PFU/cell), 10.12% of infected cells could be distinguished from the uninfected cells. These mAbs will be useful in diagnostic test development and pathogenesis studies for fish rhabdovirus. (c) 2008 Elsevier B.V. All rights reserved.; Five monoclonal antibodies (mAbs) against spring viraemia of carp (SVCV0504, isolated from common carp in China) were produced from mice immunized with purified virus preparations. The virion of SVCV contains five structural proteins, representing the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (Q. Western blotting analysis revealed that three mAbs (1145, IE10, and 11-17) recognized specifically to a single protein of 47 kDa (N), the mAb 3G4 reacted with, two SVCV0504 proteins of 69 kDa (G) and 47 kDa (N), while the mAb 1A9 reacted with three SVCV0504 proteins of 69 kDa (G), 50 kDa (P), and 47 kDa (N). By indirect ELISA, two mAbs (1H5 and 11-17) showed cross-reactivity with pike fry rhabdovirus (PFRV), but no cross-reactions with the Siniperca chuatsi rhabdovirus (SCRV), Scophthalmus maximus rhabdovirus (SMRV), Paralichthys olivaceus rhabdovirus (PoRV) were demonstrated with the five mAbs. Indirect immunofluorescence showed intense fluorescence in the cytoplasm of the SVCV0504-infected epithelioma papulosum cyprini (EPC) cells in areas corresponding to the location of granular structures. The sucrose gradient-purified SVCV0504 particles could be detected successfully by these mAbs using immunodot blotting. mAb 1A9 could completely neutralize 100 TCID50 (50% tissue culture infective dose) of SVCV0504 at a dilution of 1:8. This is the first report of development of the neutralizing mAbs against SVCV. The mAb 1A9 was analyzed further and could be used to successfully detect viral antigens in the infected-EPC cell cultures or in cryosections from experimentally infected crucian carp (Carassius auratus) by immunohistochemistry assay. Furthermore, a flow cytometry procedure for the detection and quantification of cytoplasmic SVCV0504 in cell cultures was developed with mAb 1A9. At 28 h after inoculation with the virus (0.01 PFU/cell), 10.12% of infected cells could be distinguished from the uninfected cells. These mAbs will be useful in diagnostic test development and pathogenesis studies for fish rhabdovirus. (c) 2008 Elsevier B.V. All rights reserved.
SubtypeArticle
KeywordMonoclonal Antibodies Fish Rhabdovirus Spring Viraemia Of Carp Virus (Svcv0504) Flow Cytometry
Department[Chen, Zhong-Yuan; Li, Zheng-Qiu; Zhang, Qi-Ya] Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China; [Liu, Hong] Shenzhen Exit & Entry Inspect & Quarantine Bur, Key Lab Aquat Anim Dis, Shenzhen 518001, Peoples R China
Subject AreaImmunology ; Veterinary Sciences
DOI10.1016/j.vetimm.2008.02.011
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaImmunology ; Veterinary Sciences
WOS SubjectImmunology ; Veterinary Sciences
WOS IDWOS:000256747800011
WOS KeywordHEMATOPOIETIC NECROSIS VIRUS ; SCOPHTHALMUS-MAXIMUS RHABDOVIRUS ; FLOUNDER PARALICHTHYS-OLIVACEUS ; CYPRINUS-CARPIO ; COMMON CARP ; FLOW-CYTOMETRY ; GLYCOPROTEIN GENE ; NORTH-AMERICA ; 1ST REPORT ; FISH
Citation statistics
Cited Times:28[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8090
Collection期刊论文
Corresponding AuthorZhang, QY, Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
Affiliation1.Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
2.Shenzhen Exit & Entry Inspect & Quarantine Bur, Key Lab Aquat Anim Dis, Shenzhen 518001, Peoples R China
Recommended Citation
GB/T 7714
Chen, Zhong-Yuan,Liu, Hong,Li, Zheng-Qiu,et al. Development and characterization of monoclonal antibodies to spring viraemia of carp virus[J]. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,2008,123(3-4):266-276.
APA Chen, Zhong-Yuan,Liu, Hong,Li, Zheng-Qiu,Zhang, Qi-Ya,&Zhang, QY, Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China.(2008).Development and characterization of monoclonal antibodies to spring viraemia of carp virus.VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,123(3-4),266-276.
MLA Chen, Zhong-Yuan,et al."Development and characterization of monoclonal antibodies to spring viraemia of carp virus".VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 123.3-4(2008):266-276.
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