Comparison of Indirect Immunofluorescence (IF) Test with Enzyme-Linked Immunosorbent Assay (ELISA) in Screening of Hybridomas to Very Virulent Marek's Disease Virus

For identifying virus-specific antigens of Marek's disease virus (MDV), monoclonal antibodies (MAbs) against strain Md5 of serotype 1, which is known to be a very virulent MDV (vvMDV), were isolated. Fifty-eight hybridoma clones that secreted MAbs against vvMDV were obtained. Of these MAbs, 36...

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Published in:MICROBIOLOGY and IMMUNOLOGY 1992, Vol.36(2), pp.191-197
Main Authors: Nakajima, Kazuhiro, Shibayama, Takeshi, Naito, Matao, Kurimura, Takashi, Hirai, Kanji
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - biosynthesis
Antigens, Viral - immunology
Biological and medical sciences
Enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Fluorescent Antibody Technique
Fundamental and applied biological sciences. Psychology
Herpesvirus 2, Gallid - immunology
Hybridomas - immunology
Marek's disease herpesvirus
Mice
Mice, Inbred BALB C
Microbiology
Techniques used in virology
Virology
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Summary:For identifying virus-specific antigens of Marek's disease virus (MDV), monoclonal antibodies (MAbs) against strain Md5 of serotype 1, which is known to be a very virulent MDV (vvMDV), were isolated. Fifty-eight hybridoma clones that secreted MAbs against vvMDV were obtained. Of these MAbs, 36 gave positive reactions in an immunofluorescence (IF) test, and 22 gave positive reactions on enzyme-linked immunosorbent assay (ELISA). None of these MAbs gave positive reactions in both the IF test and ELISA. Of the MAbs that gave positive reactions in the IF test, 33 clones reacted with MDV1-specific epitopes, the other three reacting with MDV1-HVT intertypic epitopes. None of the clones reacted with MDV1-MDV2 intertypic epitopes. Three virus-specific polypeptides were identified by radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) or immunoblotting. These polypeptides were recognized by 12 MAbs giving positive reactions by IF, but by none of those giving positive reactions by ELISA. In addition, size heterogeneity of the MDV1-specific phosphorylated polypeptides in the MDV1 strains was shown using the MAbs against Md5.
ISSN:0385-5600
1348-0421
DOI:10.1111/j.1348-0421.1992.tb01656.x