Abstract |
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.
|
Authors | María Soledad Serena, Christoph Geisler, Germán Ernesto Metz, Santiago Gerardo Corva, Eduardo Carlos Mórtola, Alejandra Larsen, Donald L Jarvis, María Gabriela Echeverría |
Journal | Protein expression and purification
(Protein Expr Purif)
Vol. 90
Issue 1
Pg. 1-8
(Jul 2013)
ISSN: 1096-0279 [Electronic] United States |
PMID | 23631926
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
|
Copyright | Copyright © 2013. Published by Elsevier Inc. |
Chemical References |
- Antibodies, Viral
- Recombinant Proteins
- Viral Envelope Proteins
- glycoprotein E, Suid herpesvirus 1
|
Topics |
- Animals
- Antibodies, Viral
(blood)
- Baculoviridae
(genetics)
- Enzyme-Linked Immunosorbent Assay
- Herpesvirus 1, Suid
(isolation & purification)
- Male
- Mice
- Mice, Inbred BALB C
- Pseudorabies
(diagnosis, virology)
- Recombinant Proteins
(genetics, isolation & purification, metabolism)
- Sensitivity and Specificity
- Swine
- Viral Envelope Proteins
(genetics, isolation & purification, metabolism)
|