ABSTRACT
An enzyme-linked immunonosorbent assay was established for detection of antibodies to Akabane virus in bovine sera. The assay was shown to be a useful serological tool for studies on Akabane virus infection.
Subject(s)
Antibodies, Viral/analysis , Antigens, Viral/immunology , Bunyaviridae/isolation & purification , Simbu virus/isolation & purification , Animals , Bunyaviridae Infections/diagnosis , Bunyaviridae Infections/veterinary , Cattle , Enzyme-Linked Immunosorbent Assay/veterinary , Hemagglutination Inhibition Tests/veterinary , Neutralization Tests/veterinary , Simbu virus/immunologyABSTRACT
Mouse myeloma cells (SP2/O) were fused with spleen cells from BALB/c mice immunized with detergent-solubilized antigen of purified virus, and 21 monoclonal (MC) antibodies reactive in enzyme-linked immunosorbent assay with the TO-163 strain of porcine transmissible gastroenteritis (TGE) virus were obtained. Of these MC antibodies, 14, 6 and 1 were IgG1, IgG2a and IgM, respectively. All of the MC antibodies contained light chains of the kappa type. Of these MC antibodies, 8 were found to have neutralization (NT) activity against the TO-163 strain. Comparison of 7 strains of TGE virus by NT tests using our panel of MC antibodies confirmed their close antigenic relationships, but also revealed the occurrence of distinct antigenic differences. These results suggest that there may be at least 6 different epitopes involved in NT reaction on the virion of the TO-163 strain. This notion was confirmed by the competitive binding assay.
Subject(s)
Antibodies, Monoclonal/immunology , Antigenic Variation , Antigens, Viral/immunology , Coronaviridae/immunology , Transmissible gastroenteritis virus/immunology , Animals , Antigens, Viral/analysis , Binding, Competitive , Cell Line , Enzyme-Linked Immunosorbent Assay , Epitopes , Hybridomas , Neutralization Tests , Specific Pathogen-Free Organisms , SwineABSTRACT
An enzyme-linked immunosorbent assay (ELISA) using a detergent-solubilized antigen of purified virus was developed for detection of antibody against porcine transmissible gastroenteritis (TGE) virus in swine serum. The ELISA demonstrated antibody responses in pigs immunized intramuscularly with the attenuated TO-163 strain of TGE virus and in pigs orally infected with the virulent Shizuoka strain of the virus. The results of the ELISA were well correlated with those of the neutralization test. These results indicate the usefulness of the ELISA as a serological tool for TGE virus antibody.
Subject(s)
Antibodies, Viral/analysis , Coronaviridae/immunology , Gastroenteritis, Transmissible, of Swine/immunology , Transmissible gastroenteritis virus/immunology , Animals , Enzyme-Linked Immunosorbent Assay , Immunization/veterinary , Neutralization Tests , SwineABSTRACT
ESK cells, a stable cell line derived from a swine embryo kidney, were found to be a good medium for plaque formation of the Prague and Miami strains of equine influenza virus. Factors influencing the plaque formation were investigated and a plaque assay for these viruses was worked out. The method is not only simple enough for routine use, but also is as sensitive as the egg inoculation method. The method was readily adapted for a neutralization test.
