title: The use of an in vitro microneutralization assay to evaluate the potential of recombinant VP5 protein as an antigen for vaccinating against Grass carp reovirus.
abstract: BACKGROUND
  Grass carp reovirus (GCRV) is the causative pathogen of grass carp hemorrhagic disease, one of the major diseases damaging grass carp Ctenopharyngon idellus breeding industry in China. Prevention and control of the disease is impeded largely due to the lack of research in economic subunit vaccine development. This study aimed to evaluate the potential of viral outer shell protein VP5 as subunit vaccine.
  
  
  METHODS
  The vp5 gene was isolated from the viral genome through RT-PCR and genetically engineered to express the recombinant VP5 protein in E coli. The viral origin of the recombinant protein was confirmed by Western blot analysis with a monoclonal antibody against viral VP5 protein. Polyclonal antibody against the recombinant VP5 protein was prepared from mice. A microneutralization assay was developed to test its neutralizing ability against GCRV infection in cell culture.
  
  
  RESULTS
  The GST-VP5 fusion protein (rVP5) was produced from E. Coli with expected molecular weight of 90 kDa. The protein was purified and employed to prepare anti-VP5 polyclonal antibody from mice. The anti-VP5 antibody was found to neutralize GCRV through in vitro microneutralization assay and viral progeny quantification analysis.
  
  
  CONCLUSIONS
  The present study showed that the viral VP5 protein was involved in viral infection and bacterially-expressed VP5 could be suitable for developing subunit vaccine for the control of GCRV infection.
authors.affiliation: Key Laboratory of Aquatic Genetic Resources and Utilization/Ministry of Agriculture, Shanghai Ocean University, 201306 Shanghai, China.
authors.forename: Yongxing
authors.initials: Y
authors.lastname: He
authors.forename: Hongxu
authors.initials: H
authors.lastname: Xu
authors.forename: Qian
authors.initials: Q
authors.lastname: Yang
authors.forename: Dan
authors.initials: D
authors.lastname: Xu
authors.forename: Liqun
authors.initials: L
authors.lastname: Lu
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D003530:Cyprinidae; D005260:Female; D005393:Fish Diseases / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D012087:Reoviridae / Q000235:genetics / Q000276:immunology*; D012088:Reoviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary* / Q000821:virology; D014611:Vaccination; D014764:Viral Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D014765:Viral Vaccines