Agar gel immunodiffusion analysis using baculovirus-expressed recombinant bovine leukemia virus envelope glycoprotein (gp51/gp30(T-)).
Bovine leukemia virus (BLV) envelope glycoprotein (gp51/ gp30(T-)), consisting of BLV gp51 and BLV gp30 that lacked its C-terminal transmembrane domain, was expressed in insect cells under the control of the baculovirus polyhedron promoter. Recombinant BLV gp51/gp30(T-) secreted from insect cells was determined by immunofluorescence, enzyme-linked immunosorbent and western blot assays using a BLV-specific monoclonal antibody and BLV-positive bovine antibodies. An agar gel immunodiffusion (AGID) test using gp51/gp30(T-) as the antigen for the detection of BLV antibodies in serum was developed and compared to traditional AGID, which uses wild type BLV antigen derived from fetal lamb kidney cells. AGID with the recombinant BLV gp51/gp30(T-) was relatively more sensitive than traditional AGID. When the two methods were tested with bovine sera from the field, the recombinant BLV gp51/gp30(T-) and traditional antigen had a relative sensitivity of 69.8% and 67.4%, respectively, and a relative specificity of 93.3% and 92.3%. These results indicated that the recombinant BLV gp51/gp30(T-) is an effective alternative antigen for the diagnosis of BLV infection in cattle.
Lim
Seong In
SI
National Veterinary Research and Quarantine Service, Anyang 430-757, Korea.
Jeong
Wooseog
W
Tark
Dong Seob
DS
Yang
Dong Kun
DK
Kweon
Chang Hee
CH
D000362:Agar; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D016583:Enzootic Bovine Leukosis / Q000097:blood / Q000276:immunology; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D005779:Immunodiffusion / Q000379:methods / Q000662:veterinary*; D007668:Kidney / Q000166:cytology; D001909:Leukemia Virus, Bovine / Q000235:genetics / Q000378:metabolism*; D008967:Molecular Biology; D012756:Sheep; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins; D000362:Agar