Production of pseudorabies virus recombinant glycoprotein B and its use in an agar gel immunodiffusion (AGID) test for detection of antibodies with sensitivity and specificity equal to the virus neutralization assay.

MARÍA SOLEDAD SERENA; CHRISTOPH GEISLER; GERMÁN ERNESTO METZ; EDUARDO CARLOS MÓRTOLA; MARÍA GABRIELA ECHEVERRÍA

JOURNAL OF VIROLOGICAL METHODS

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2016 vol. 230 p. 9 - 12

0166-0934

Pseudorabies virus (PrV) causes Aujeszky?s disease (AD), which affects mainly swine, but also cattle, sheep,and wild animals, resulting in substantial economic losses due to animal mortality and lost productivityworldwide. To combat PrV, eradication programs using PrV strains lacking the gene encoding glycoproteinE (gE) are ongoing in several countries. These eradication programs have generated a currently unmetdemand for affordable, easy-to-use, and sensitive tests that can detect PrV infection in pigs infected witheither wild-type virus or vaccine strain (gE-deleted) virus. To meet this demand, we used the baculovirusinsectcell system to produce recombinant glycoprotein B (gB) as antigen for an immune assay. The highGC-content (70% average) of the gB gene from the Argentinian PrV CL15 strain necessitated the use ofbetaine as a PCR enhancer to amplify the extracellular domain. Recombinant gB was expressed at highlevels and reacted strongly with sera from PrV infected pigs. We used the recombinant gB to developan agar gel immunodiffusion (AGID) test for detection of PrV antibodies. Compared to the gold standardvirus neutralization (VN) assay, the AGID sensitivity and specificity were 95% and 96.6% respectively.Thus, recombinant gB produced in the baculovirus-insect cell system is a viable source of antigen forthe detection of PrV antibodies in AGID tests. Considering its relatively lower cost, simplicity of use andresult interpretation, our AGID is a valuable alternative tool to the VN assay.