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Clinical and Diagnostic Laboratory Immunology, January 2004, p. 147-151, Vol. 11, No. 1
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.1.147-151.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Expression of the Bovine Leukemia Virus Envelope Glycoprotein (gp51) by Recombinant Baculovirus and Its Use in an Enzyme-Linked Immunosorbent Assay

Antonio De Giuseppe, Francesco Feliziani, Domenico Rutili, and Gian Mario De Mia*

Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, 06126 Perugia, Italy

Received 30 May 2003/ Returned for modification 14 September 2003/ Accepted 21 October 2003

The gene encoding the major envelope glycoprotein (gp51) with its signal sequence, represented by an additional NH2-terminal 33-residue amino acid sequence of bovine leukemia virus (BLV), was inserted into a baculovirus transfer vector. A recombinant virus expressing a secreted gp51 protein in insect cells was isolated. The recombinant gp51 expressed was characterized by using an anti-BLV monoclonal antibody by both Western blotting analysis and enzyme-linked immunosorbent assay (ELISA). The secreted gp51 was used as an antigen, and an ELISA with recombinant gp51 (rgp51) was developed for the detection of BLV antibodies. This new procedure was compared with a previous ELISA method for the detection of BLV antibodies and an agar gel immunodiffusion test performed with an unpurified BLV antigen preparation. The comparative testing of field samples showed that the ELISA with rgp51 is more specific and also suitable for the testing of pooled sera.

* Corresponding author. Mailing address: Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, via G. Salvemini 1, 06126 Perugia, Italy. E-mail: gm.demia{at}izs.it.

Clinical and Diagnostic Laboratory Immunology, January 2004, p. 147-151, Vol. 11, No. 1
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.1.147-151.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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Copyright © 2004 by the American Society for Microbiology. All rights reserved.