Preparation and Specificity Identification of Monoclonal Antibody against Serotype 1 Marek's Disease Virus
To prepare the monoclonal antibody(McAb)of serotype 1 Marek's disease virus,BALB/c mice were immunized with the serotype 1 MDV CVI988/Rispens strain concentrated by ultracentrifugation,and McAbs were prepared by hybridoma technique.By subcloning,indirect ELISA and indirect immunofluorescence assay(IFA)identification,three positive McAb cell lines were obtained and named as 4D9,1C5 and 1F10,respectively.Then,the specificity of the obtained McAbs was analyzed.The ELISA titers of 4D9,1C5 and 1F10 reached 5·1×104,8.2×105 and 4.1×105,indicating that all of the three could be used to establish ELISA detection methods;IFA results showed that the ascites prepared by 4D9,1C5 and 1F10 had good reactivity with different serotype 1 MDV strains with the titers of 1∶12800,1∶25600 and 1∶12800,respectively,and all of the three have no cross-reactivity with serotype 3 MDV strains and other common avian-derived viruses;The subtype test of the three above McAbs indicated that 4D9 was IgG2a/к subtype,1C5 was IgG2b/κ subtype and 1F10 was IgG1/κ subtype.The IFA method established with 1C5 could detect more than 5 PFU CVI988/Rispens,confirming that this method is suitable for the detection of serotype 1 MDV.In summary,the McAbs obtained in this study can be used for the detection of serotype 1 MDV and provide the basis for further researches on MDV pathogenicity and the development of diagnostic reagents.
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