A highly specific and sensitive competitive enzyme-linked immunosorbent assay (ELISA) based on baculovirus expressed pseudorabies virus glycoprotein gE and gI complex.
A direct competition enzyme-linked immunosorbent assay (ELISA) based on baculovirus expressed complex of pseudorabies virus (PRV) glycoproteins E (gE) and I (gI) has been developed. For that purpose gE and gI genes of PRV were co-expressed in insect cells. Complex formation was confirmed by radioimmunoprecipitation assay. The specificity and sensitivity of the test were evaluated and compared with an ELISA using only gE as an antigen and a commercially available test. For validation, 245 negative sera and 165 positive sera have been tested. The gE/gI ELISA had a higher sensitivity and specificity when compared with the ELISA using only gE as the antigen. Both sensitivity and specificity were comparable with the commercially available test. Moreover, the test based on the baculovirus gE/gI complex allows the detection of anti-gE antibodies in pig serum as early as two weeks after infection. The gE/gI ELISA test is easy to perform; its additional advantage is that the gE/gI antigen can be produced in baculovirus system in large quantities without handling live pseudorabies virus.
Gut
M
M
University of Gdansk, Department of Molecular Virology, Poland. monia@biotech.univ.gda.pl
Jacobs
L
L
Tyborowska
J
J
Szewczyk
B
B
Bienkowska-Szewczyk
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D003124:Colorimetry / Q000662:veterinary; D004279:DNA, Viral / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D011558:Herpesvirus 1, Suid / Q000276:immunology*; D011557:Pseudorabies / Q000175:diagnosis* / Q000276:immunology; D015531:Radioimmunoprecipitation Assay / Q000662:veterinary; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D018411:Spodoptera; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000276:immunology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral; D014759:Viral Envelope Proteins; C089580:glycoprotein E, Suid herpesvirus 1; C042966:pseudorabies virus glycoproteins