TY - JOUR
T1 - A highly specific and sensitive sandwich blocking ELISA based on baculovirus expressed pseudorabies virus glycoprotein B
AU - Gut-Winiarska, Monika
AU - Jacobs, Liesbeth
AU - Kerstens, Hinri
AU - Bienkowska-Szewczyk, Krystyna
N1 - Funding Information:
This work was supported by Polish KBN grant No. 1085/PO4/97/12. M. Gut, while working in ID-Lelystad, was supported by MOE project No. 47301-100.
PY - 2000/7
Y1 - 2000/7
N2 - 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. (C) 2000 Elsevier Science B.V.
AB - 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. (C) 2000 Elsevier Science B.V.
KW - Aujeszky's disease
KW - Baculovirus expression
KW - Blocking ELISA
KW - Glycoprotein B
UR - http://www.scopus.com/inward/record.url?scp=0033879538&partnerID=8YFLogxK
U2 - 10.1016/S0166-0934(00)00178-6
DO - 10.1016/S0166-0934(00)00178-6
M3 - Article
C2 - 10921843
AN - SCOPUS:0033879538
SN - 0166-0934
VL - 88
SP - 63
EP - 71
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1
ER -