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.
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: Department of Virology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Argentina.
authors: María Soledad
authors: MS
authors: Serena
authors: Christoph
authors: C
authors: Geisler
authors: Germán Ernesto
authors: GE
authors: Metz
authors: Santiago Gerardo
authors: SG
authors: Corva
authors: Eduardo Carlos
authors: EC
authors: Mórtola
authors: Alejandra
authors: A
authors: Larsen
authors: Donald L
authors: DL
authors: Jarvis
authors: María Gabriela
authors: MG
authors: Echeverría
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D011558:Herpesvirus 1, Suid / Q000302:isolation & purification*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011557:Pseudorabies / Q000175:diagnosis* / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C089580:glycoprotein E, Suid herpesvirus 1
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