Preparation of serotype-Ⅲ specific monoclonal antibody against MDV and its pre-liminary applications in plaque counting of vaccines
In order to prepare and characterize serotype-Ⅲ specific monoclonal antibody(McAb)a-gainst Marek's disease virus(MDV),and then utilized it for establishing plaque counting method by indirect immunofluorescent staining.Purified MDV serotype-Ⅲ HVT Fc-126 strains were used to immunize BALB/c mice.The hybridoma cultured supernatants were screened by indirect immu-nofluorescent assay with HVT Fc-126 infected chicken embryo fibroblast cells(CEF).The hybri-doma clones were selected and employed for preparation of McAb.Plaque counting method by indi-rect immunofluorescent staining was established with the McAb to detect the MDV serotype-Ⅲplaque forming units(PFU)in Marek's Disease bivalent live vaccines.Two hybridoma clones that constitutively secrete McAb against MDV serotype-Ⅲ were screened.High purified McAb was pre-pared with 1D2 hybridoma clone.Compared with agarose plaque assay,the indirect immunofluores-cent staining method characterized higher sensitivity,specificity,and more rapidity.Significantly,the indirect immunofluorescent staining method could accurately count PFU of serotype-Ⅲ viruses in bivalent vaccine containing two serotype viruses.Hence,serotype-Ⅲ specific McAb against MDV has been prepared successfully,and could be applied in plaque counting of bivalent vaccines by in-direct immunofluorescent staining method.
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