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Author Tin, C.M.; Yuan, L.; Dexter, R.J.; Parra, G.I.; Bui, T.; Green, K.Y.; Sosnovtsev, S.V. doi  openurl
  Title (up) A Luciferase Immunoprecipitation System (LIPS) assay for profiling human norovirus antibodies Type Research Support, N.I.H., Intramural
  Year 2017 Publication Journal of virological methods Abbreviated Journal J Virol Methods  
  Volume 248 Issue Pages 116-129  
  Corporate Author Thesis  
  Address Caliciviruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, DHHS, Bethesda, MD, USA. Electronic address:  
  Keywords Animals; Antibodies, Viral/*blood/*immunology/isolation & purification; Antibody Specificity; Caliciviridae Infections/immunology/virology; Capsid Proteins/genetics/*immunology; Cross Reactions; Enzyme-Linked Immunosorbent Assay/methods; Epitopes; Humans; Immunoprecipitation/*methods; Luciferases, Renilla/genetics/*immunology; Norovirus/*immunology/isolation & purification; Recombinant Fusion Proteins/immunology; Swine; Swine, Miniature; Vaccines, Virus-Like Particle/immunology; Viral Vaccines/immunology; *Elisa; *Human norovirus; *LIPS assay; *Serum antibody  
  Abstract A luciferase immunoprecipitation systems (LIPS) assay was developed to define the antigenic specificity and titer of antibodies directed against human norovirus (HuNoV). Recombinant proteins, expressed by plasmid constructs encoding Renilla luciferase (Ruc) fused to the full-length HuNoV major capsid protein (VP1) (Ruc-antigen), were generated for ten HuNoV strains. In addition, subdomain constructs Ruc-Shell (S) and Ruc-Protruding (P) were engineered for a representative GII.4 norovirus (strain GII.4/2006b). The LIPS assay measured antibody levels in a well-defined panel of HuNoV-specific sera, and the results were compared to an ELISA standard. In hyperimmune sera, the LIPS produced titers similar to or higher than those measured by the ELISA of HuNoV-specific antibodies. The specificity of antibodies in various sera was profiled by LIPS with a panel of diverse Ruc-antigens containing full-length HuNoV VP1 proteins or VP1 subdomains, and the assay detected both specific and cross-reactive antibodies. Competition assays, in which antibodies were pre-incubated with one or more intact VLPs representing different genotypes, proved useful in further assessment of the antibody specificity detected by LIPS in complex polyclonal sera. The profiling of HuNoV-specific antibodies in the high-throughput LIPS format may prove useful in defining the strength or specificity of the adaptive immune response following natural infection or vaccination.  
  Publisher Place of Publication Editor  
  Language English Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 0166-0934 ISBN Medium  
  Area Expedition Conference  
  Notes PMID:28673856 Approved no  
  Call Number NCSU @ edshirle @ Serial 3649  
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