Expression of non-structural proteins of Senecavirus A and preparation of the polyclonal antibodies
In order to differentiate and diagnose whether pigs were infected with Senecavirus A(SVA),recombinant plasmids expressing SVA 2C,3AB,3C,and 3D1(1-160aa)were constructed through prokar-yotic expression,respectively.The purified recombinational proteins were used to immunize Rex rabbits to prepare polyclonal antibodies.Immunogenicity was evaluated using Western-blot and IFA,and serum antibody titers were detected using ELISA.Western-blot analysis showed that the prepared polyclonal antibodies could react specifically with the corresponding recombinant proteins SVA 2C,3AB,3C,or 3D1.The ELISA titers of SVA 2C,3AB,3C,and 3D1 polyclonal antibodies were 1∶1.1×1018,1∶1.67×1010,1∶2.1×1012,and 1∶4.4×1015,respectively.IFA showed that the polyclonal antibodies could specifically recognize SVA viral proteins.In summary,this study successfully expressed four nonstruc-tural proteins of SVA and prepared polyclonal antibodies with good immunoreactivity,which laid the foundation for the differential diagnosis of SVA infection.
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