Agar gel immunodiffusion analysis using baculovirus-expressed recombinant bovine leukemia virus envelope glycoprotein (gp51/gp30T-).
10.4142/jvs.2009.10.4.331
- Author:
Seong In LIM
1
;
Wooseog JEONG
;
Dong Seob TARK
;
Dong Kun YANG
;
Chang Hee KWEON
Author Information
1. National Veterinary Research and Quarantine Service, Anyang 430-757, Korea. kweonch@mail.nvrqs.go.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
AGID;
baculovirus expression;
bovine leukemia virus;
glycoproteins
- MeSH:
Agar;
Animals;
Antibodies, Viral/blood;
Antigens, Viral/immunology;
Baculoviridae/*metabolism;
Cattle;
Cell Line;
Enzootic Bovine Leukosis/blood/immunology;
Gene Expression Regulation, Viral/*physiology;
Immunodiffusion/methods/*veterinary;
Kidney/cytology;
Leukemia Virus, Bovine/genetics/*metabolism;
Molecular Biology;
Sheep;
Viral Envelope Proteins/genetics/*metabolism
- From:Journal of Veterinary Science
2009;10(4):331-336
- CountryRepublic of Korea
- Language:English
-
Abstract:
Bovine leukemia virus (BLV) envelope glycoprotein (gp51/gp30T-), 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/gp30T- 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/gp30T- 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/gp30T- was relatively more sensitive than traditional AGID. When the two methods were tested with bovine sera from the field, the recombinant BLV gp51/gp30T- 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/gp30T- is an effective alternative antigen for the diagnosis of BLV infection in cattle.
