Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/293560
COMPARTIR / EXPORTAR:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Título: | DIVA diagnostic of Aujeszky's disease using an insect-derived virus glycoprotein E |
Autor: | Gómez-Sebastián, S.; Pérez-Filgueira, M.; Gómez Casado, Eduardo; Nuñez, M. del Carmen; Sánchez-Ramos, Ismael CSIC ORCID; Tabarés, E.; Escribano, J. M. | Palabras clave: | PRV gE glycoprotein expression Baculovirus Larvae Low-cost Insect expression system |
Fecha de publicación: | 2008 | Editor: | Elsevier | Citación: | Journal of Virological Methods 153(1): 29-35 (2008) | Resumen: | Commercial vaccines against Aujeszky's disease are mainly formulated using deleted versions of attenuated or inactivated Pseudorabies virus (PRV) particles lacking of the structural glycoprotein E (gE). Complementary diagnostic assays used to differentiate infected from vaccinated animals (DIVAs), are based on the detection of serum antibodies against gE. A recombinant version of the PRV gE protein was expressed in a baculovirus vector system in Trichoplusia ni insect larvae in order to obtain this diagnostic reagent for large scale diagnosis at reduced costs. A recombinant gE gene (gEr), lacking of signal peptide and transmembrane domains, was cloned into a modified baculovirus vector to allow glycosylation of the protein and its subsequent exportation to the extracellular space. Analysis by SDS-PAGE, Western-blotting and glycoprotein staining revealed that a glycosylated protein of the expected electrophoretic mobility was obtained in infected larvae. Time course experiments revealed that maximum expression levels were reached 72 h post-infection using 104 pfu of the recombinant baculovirus (BACgEr) per inoculated larva. An indirect PRV gE-ELISA was developed using gEr as a coating antigen. A comparison between larvae-derived PRV gE-ELISA and two commercially available PRV diagnostic kits showed good correlation between assays and better sensitivity when testing certain sera pig samples using the gEr ELISA. More than 30,000 ELISA determinations could be performed from crude extracts obtained from a single larva infected with the recombinant baculovirus, indicating the feasibility of this strategy for inexpensive production of glycosylated antigens for PRV diagnosis. © 2008 Elsevier B.V. All rights reserved. | URI: | http://hdl.handle.net/10261/293560 | DOI: | 10.1016/j.jviromet.2008.06.017 | ISSN: | 0166-0934 |
Aparece en las colecciones: | (INIA) Artículos |
Mostrar el registro completo
CORE Recommender
SCOPUSTM
Citations
27
checked on 13-may-2024
WEB OF SCIENCETM
Citations
25
checked on 23-feb-2024
Page view(s)
24
checked on 16-may-2024
Google ScholarTM
Check
Altmetric
Altmetric
NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.