Noninfectious virus-like particle antigen for detection of swine vesicular disease virus antibodies in pigs by enzyme-linked immunosorbent assay.
An inactivated SVDV antigen is used in current enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies to swine vesicular disease virus (SVDV). To develop a noninfectious recombinant alternative, we produced SVDV-like particles (VLPs) morphologically and antigenically resembling authentic SVDV particles by using a dual baculovirus recombinant, which expresses simultaneously the P1 and 3CD protein genes of SVDV under different promoters. Antigenic differences between recombinant VLPs and SVDV particles were not statistically significant in results obtained with a 5B7-ELISA kit, indicating that the VLPs could be used in the place of SVDV antigen in ELISA kits. We developed a blocking ELISA using the VLPs and SVDV-specific neutralizing monoclonal antibody 3H10 (VLP-ELISA) for detection of SVDV serum antibodies in pigs. The VLP-ELISA showed a high specificity of 99.9% when tested with pig sera that are negative for SVDV neutralization (n=1,041). When tested using sera (n=186) collected periodically from pigs (n=19) with experimental infection with each of three different strains of SVDV, the VLP-ELISA detected SVDV serum antibodies as early as 3 days postinfection and continued to detect the antibodies from all infected pigs until termination of the experiments (up to 121 days postinfection). This test performance was similar to that of the gold standard virus neutralization test and indicates that the VLP-ELISA is a highly specific and sensitive method for the detection of SVDV serum antibodies in pigs. This is the first report of the production and diagnostic application of recombinant VLPs of SVDV. Further potential uses of the VLPs are discussed.
Ko
Young-Joon
YJ
National Veterinary Research and Quarantine Service, 480 Anyang-6 dong, Anyang, Kyong-gi 430-824, Korea.
Choi
Kang-Seuk
KS
Nah
Jin-Ju
JJ
Paton
David J
DJ
Oem
Jae-Ku
JK
Wilsden
Ginette
G
Kang
Shien-Young
SY
Jo
Nam-In
NI
Lee
Joo-Ho
JH
Kim
Jae-Hong
JH
Lee
Hee-Woo
HW
Park
Jong-Myeong
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000302:isolation & purification*; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D007313:Insecta / Q000166:cytology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D013552:Swine; D013555:Swine Vesicular Disease / Q000097:blood / Q000175:diagnosis* / Q000821:virology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins