A highly specific and sensitive sandwich blocking ELISA based on baculovirus expressed pseudorabies virus glycoprotein B.
A direct sandwich blocking enzyme-linked immunosorbent assay (BacgB ELISA) based on the reaction between a monoclonal antibody (MAb) and a recombinant glycoprotein B (gB) of pseudorabies virus (PRV) was developed. This protein was obtained in large quantities from insect cells infected with a PRV gB recombinant baculovirus. Expression of the gB was confirmed by immunoperoxidase monolayer assay (IPMA) with gB specific MAbs. The specificity and sensitivity of the developed BacgB ELISA were evaluated and compared with two commercially available tests by using sets of sera of known PRV infection or vaccination history. For validation, 347 serum samples have been tested. The BacgB ELISA had a high sensitivity and specificity, which were comparable with those of the two commercial tests. In addition, the BacgB ELISA allows detecting anti-gB antibodies in pig serum as early as 7 days following infection. Also maternal antibodies in uninfected pig sera were detected. We conclude that the BacgB ELISA is a useful tool for the detection of as well vaccinated as infected pigs (including derivatives from gE negative vaccine strains), with the added advantage that it uses an antigen that can be produced safely and in large quantities.
Department of Molecular Virology, University of Gdansk, Faculty of Biotechnology, Poland. monia@biotech.univ.gda.pl
M
M
Gut-Winiarska
L
L
Jacobs
H
H
Kerstens
K
K
Bienkowska-Szewczyk
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D011558:Herpesvirus 1, Suid / Q000276:immunology* / Q000378:metabolism; D011557:Pseudorabies / Q000175:diagnosis* / Q000821:virology; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
D000914:Antibodies, Viral; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C070915:glycoprotein gII, pseudorabies virus