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

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 nonstructu...

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Bibliographic Details
Published in:Journal of virological methods 2011-02, Vol.171 (2), p.388-393
Main Authors: Serena, María Soledad, Metz, Germán Ernesto, Corva, Santiago Gerardo, Mórtola, Eduardo Carlos, Echeverría, María Gabriela
Format: Article
Language:English
Subjects:
Animals
antibodies
Antibodies, Viral - blood
antigen detection
Aujeszky disease
Baculoviridae
Baculovirus
Biological and medical sciences
Bluetongue virus
Bovine leukemia virus
Cell Line
Diagnosis, Differential
enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Foot-and-mouth disease virus
Fundamental and applied biological sciences. Psychology
Gene Expression
genes
Genetic Vectors
Glycoprotein E
Hepatitis B virus
Herpesvirus 1, Suid - immunology
Human immunodeficiency virus 1
Immunodominant epitopes
Influenza A virus
Insecta
Microbiology
Orthomyxoviridae
polymerization
Pseudorabies - diagnosis
Pseudorabies - immunology
Pseudorabies - virology
Pseudorabies virus
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - isolation & purification
recombinant proteins
SHV-1
Suid herpesvirus 1
Swine
Techniques used in virology
ultracentrifugation
Viral Envelope Proteins - genetics
Viral Envelope Proteins - isolation & purification
viral nonstructural proteins
Viral Nonstructural Proteins - genetics
Viral Vaccines - immunology
Virology
Virology - methods
viruses
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Description
Summary: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.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2010.12.005