Development of an AGID based on baculovirus expressed Pseudorabies virus glycoprotein B

SERENA, MS; METZ, GE; PANEI CJ; LARSEN A; ASPITIA C; GALOSI C. M; MORTOLA E; ECHEVERRÍA M. G.

Medellín

Congreso; 11th Congress of the Latin American Association of Immunology (ALAI) and 10th Congress of the Asociación Colombiana de Alergia, Asma e Inmunología (ACAAI) IMMUNOCOLOMBIA 2015; 2015

The pseudorabies virus (PrV) is a member of the genus Varicellovirus, subfamily Alphaherpesvirinae and family Herpesviridae, characterized as the etiologic agent of Aujeszky?s disease in swine. Aujeszky?s disease is a one of the most infectious diseases affecting the swine industry which causes central nervous system symptoms and is characterized by an acute and often fatal infection in piglets, while in older pigs it causes disorders such as encephalitis, pneumonia and abortion. The glycoprotein B, a major constituent of the viral envelope, exists has a disulphide-linked complex consisting of three glycoproteins: gBa, gBb and gBc. The primary translation product consists of 913aa, while the mature form of the glycoprotein after remove the signal sequence has 855aa. The fully glycosylated (gBa) is a 120kDa polypeptide that is cleaved between Arg444 and Ser445 by a cellular protease to yield two subunits, gBb (68kDa) and gBc (55kDa). The gB is indispensable for virus replication, plays an essential role in membrane fusion events during virus infection and cell-to-cell spread of virus and belongs to the class of glycoproteins that are most conserved among herpesviruses. Serological tests such as virus neutralization (VN) and ELISA require special facilities or commercial equipment, in contrast, the AGID which was designed for other virus animal?s as an effective tool for the detection of specific antibodies, is a fast technique that does not require special conditions and can be performed at any veterinary diagnostic laboratory. The baculovirus expression system has been used to produce large quantities of antigenically active and processed freeing glycoproteins. This report describes the development of a simple AGID test based on the gB of PrV produced in BaculoDirect Expression System to examine a porcine serum samples with a sensitivity and specificity comparable with VN test (gold standart test).