Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs

Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycopro...

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Autores principales: Serena, María Soledad, Geisler, Christoph, Metz, Germán Ernesto, Corva, Santiago Gerardo, Mórtola, Eduardo Carlos, Larsen, Alejandra Edith, Jarvis, Donald L., Echeverría, María Gabriela
Formato: Articulo
Lenguaje:Inglés
Publicado: 2013
Materias:
Ciencias Veterinarias
Suid Herpesvirus 1
Baculovirus
Insect cell
Glycoprotein
ELISA
GC content
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/129004
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-129004
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Veterinarias
Suid Herpesvirus 1
Baculovirus
Insect cell
Glycoprotein
ELISA
GC content
spellingShingle Ciencias Veterinarias
Suid Herpesvirus 1
Baculovirus
Insect cell
Glycoprotein
ELISA
GC content
Serena, María Soledad
Geisler, Christoph
Metz, Germán Ernesto
Corva, Santiago Gerardo
Mórtola, Eduardo Carlos
Larsen, Alejandra Edith
Jarvis, Donald L.
Echeverría, María Gabriela
Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs
topic_facet Ciencias Veterinarias
Suid Herpesvirus 1
Baculovirus
Insect cell
Glycoprotein
ELISA
GC content
description Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
format Articulo
Articulo
author Serena, María Soledad
Geisler, Christoph
Metz, Germán Ernesto
Corva, Santiago Gerardo
Mórtola, Eduardo Carlos
Larsen, Alejandra Edith
Jarvis, Donald L.
Echeverría, María Gabriela
author_facet Serena, María Soledad
Geisler, Christoph
Metz, Germán Ernesto
Corva, Santiago Gerardo
Mórtola, Eduardo Carlos
Larsen, Alejandra Edith
Jarvis, Donald L.
Echeverría, María Gabriela
author_sort Serena, María Soledad
title Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs
title_short Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs
title_full Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs
title_fullStr Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs
title_full_unstemmed Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky's disease in infected pigs
title_sort expression and purification of suid herpesvirus-1 glycoprotein e in the baculovirus system and its use to diagnose aujeszky's disease in infected pigs
publishDate 2013
url http://sedici.unlp.edu.ar/handle/10915/129004
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