title: A differential ELISA based on recombinant immunodominant epitopes of the gE gene of SHV-1 in a baculovirus-insect cell system to discriminate between pigs infected naturally with pseudorabies and vaccinated pigs.
abstract: In the present study, the fragment corresponding to the immunodominant epitopes of the gE gene (gEpi) from the CL15 Argentinean strain of pseudorabies virus was expressed successfully in a baculovirus-insect cell system that contained the M6 gene of Bluetongue virus, which encodes the NS1 nonstructural protein. This protein has the ability to polymerize into highly immunogenic tubules inside infected cells that can be purified at large quantities by ultracentrifugation. Previously, the NS1 protein has been expressed by fusing it to sequences derived from viruses, such as human immunodeficiency virus type 1, hepatitis B virus, bovine leukemia virus, foot-and-mouth disease virus and influenza A virus. In the present study, a recombinant protein was obtained containing the gEpi fused to NS1 (NS1-gEpi) and used it as ELISA antigen for detection of anti-gE antibodies in order to discriminate between infected and vaccinated animals. This is the first report where gEpi was expressed in this particular baculovirus-insect cell system.
authors.affiliation: Virology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Argentina.
authors.forename: María Soledad
authors.initials: MS
authors.lastname: Serena
authors.forename: Germán Ernesto
authors.initials: GE
authors.lastname: Metz
authors.forename: Santiago Gerardo
authors.initials: SG
authors.lastname: Corva
authors.forename: Eduardo Carlos
authors.initials: EC
authors.lastname: Mórtola
authors.forename: María Gabriela
authors.initials: MG
authors.lastname: Echeverría
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D016367:Baculoviridae; D002460:Cell Line; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015870:Gene Expression; D005822:Genetic Vectors; D011558:Herpesvirus 1, Suid / Q000276:immunology*; D007313:Insecta; D011557:Pseudorabies / Q000175:diagnosis* / Q000276:immunology* / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000302:isolation & purification; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014765:Viral Vaccines / Q000276:immunology*; D014773:Virology / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; C078966:NS1 protein, bluetongue virus; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C089580:glycoprotein E, Suid herpesvirus 1