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Comparison of Indirect Immunofluorescence (IF) Test with Enzyme‐Linked Immunosorbent Assay (ELISA) in Screening of Hybridomas to Very Virulent Marek's Disease Virus
Author(s) -
Nakajima Kazuhiro,
Shibayama Takeshi,
Naito Matao,
Kurimura Takashi,
Hirai Kanji
Publication year - 1992
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1992.tb01656.x
Subject(s) - epitope , marek's disease , monoclonal antibody , virology , biology , virus , gel electrophoresis , antigen , microbiology and biotechnology , virulence , antibody , immunofluorescence , polyacrylamide gel electrophoresis , enzyme , biochemistry , gene , immunology
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.