scholarly journals Molecular Cloning, Expression, and Antigenicity of Seto Virus Belonging to Genogroup I Norwalk-Like Viruses

2000 ◽  
Vol 38 (9) ◽  
pp. 3492-3494 ◽  
Author(s):  
Shinichi Kobayashi ◽  
Kenji Sakae ◽  
Yasumoto Suzuki ◽  
Kuniko Shinozaki ◽  
Mineyuki Okada ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Capsid Protein ◽  
Immunosorbent Assay ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Viral Capsid ◽  
Specific Detection ◽  
Japanese Strain ◽  
Baculovirus Expression

The viral capsid protein of the Seto virus (SeV), a Japanese strain of genogroup I Norwalk-like viruses (NLVs), was expressed as virus-like particles using a baculovirus expression system. An antigen detection enzyme-linked immunosorbent assay based on hyperimmune antisera to recombinant SeV was highly specific to homologous SeV-like strains but not heterologous strains in stools, allowing us type-specific detection of NLVs.

DETECTION OF ANTI-RETICULOENDOTHELIOSIS ANTIBODY BY ENZYME-LINKED IMMUNOSORBENT ASSAY USING ENVELOPE PROTEIN EXPRESSED IN BACULOVIRUS

2016 ◽  
Vol 42 (03) ◽  
pp. 165-170
Author(s):  
Hui-Wen Chen ◽  
Ting-Hsiang Hsiao ◽  
Ching-Ho Wang
Keyword(s):  
Immunosorbent Assay ◽  
Envelope Protein ◽  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Relative Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Baculovirus Expression ◽  
Relative Specificity

Reticuloendotheliosis virus (REV) infects various animals including chickens, ducks, geese, pheasants, peafowl and other birds. REV causes tumor, runting disease syndrome and immunosuppression in infected birds. The purpose of this study was to develop a rapid, reliable and convenient method to detect anti-REV antibody. The REV envelope protein was expressed by baculovirus expression system and evaluated for the potential as an antigen in an enzyme linked-immunosorbent assay (ELISA). The env gene from a REV strain goose/3410/06 was cloned into bacmid vector and expressed in Sf9 insect cells. The expected recombinant envelope protein expressed in infected insect cells was demonstrated in western blot with size of 62 kDa, and purified for using as antigen in an ELISA. A total of 182 chicken serum samples were used to evaluate the suitability of the ELISA based on neutralization test as the gold standard. The results showed that the relative sensitivity and relative specificity of this ELISA were 88.5% and 97.7%, respectively. The present ELISA using baculovirus expression envelope protein can be used to detect anti-REV antibody in the field.

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