Development of an enzyme-linked immunosorbent assay using recombinant chicken anemia virus proteins expressed in a baculovirus vector system.
Recombinant baculoviruses were constructed to express the putative proteins VP1, VP2 or VP3 of the chicken anemia virus (CAV). The recombinant VP1, VP2 or VP3 were detected by SDS-PAGE, and their molecular weights were 50, 30/27 and 16 kDa, respectively. The VP2 and VP3 reacted with sera from CAV-infected chickens in Western blot analysis and when used as an enzyme-linked immunosorbent assay (ELISA) antigen, but VP1 did not. Antibodies to CAV were detected, by ELISA using crude insect cell lysates containing VP2 or VP3, from 2 to 20 weeks or 2 to 7 weeks after CAV infection, respectively. These findings indicate that recombinant VP2 and VP3 expressed in the baculovirus vector system can be used as antigens to detect anti-CAV antibodies in ELISA.
Iwata
N
N
Nippon Institute for Biological Science, Tokyo, Japan.
Fujino
M
M
Tuchiya
K
K
Iwata
A
A
Otaki
Y
Y
Ueda
S
S
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000917:Antibody Formation; D000956:Antigens, Viral / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D016367:Baculoviridae; D015153:Blotting, Western; D002460:Cell Line; D017727:Chicken anemia virus / Q000276:immunology / Q000302:isolation & purification*; D002645:Chickens; D018173:Circoviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007564:Japan; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology; D018411:Spodoptera; D013601:T-Lymphocytes; D014162:Transfection; D014764:Viral Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins