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Swine Acute Diarrhea Syndrome Coronavirus (SADS-CoV) is a member of the Coronaviridae family. The virus is associated with severe small intestine inflammation and diarrhea in suckling piglets. In 2017, SADS-CoV was first detected and identified as the causative agent of a devasting swine disease outbreak in southern China. Routine monitoring and early detection of the source of infection is therefore integral to the prevention and control of a SADS-CoV outbreak in the United States. However, the United States does not currently have any diagnostic or surveillance tests to identify this emerging disease. To address these industry needs, we developed monoclonal antibodies against SADS-CoV. To start with, the nonstructural SADS-CoV, papain-like protease 2 region (PLP2) was cloned into an E.coli plasmid expression vector Next, the nonstructural protein SADS-CoV PLP2 was expressed, purified and used as antigen. Then, the purified antigen was used to immunized mice to produce monoclonal antibodies and rabbits were immunized to produce antisera containing polyclonal antibodies also for diagnostic test development. Mouse spleen cells and a mouse tumor cell line (NS-1 myeloma) were fused together to produce hybridoma clones. The resulting monoclonal antibodies were characterized and evaluated for their ability to specifically recognize SADS-CoV PLP2 epitope in cell culture. In total, nine monoclonal antibodies and approximately 105 milliliters of rabbit antisera were produced. The immunoreactivity of each monoclonal antibody and antisera was tested by enzyme linked immunosorbent assay (ELISA), immunofluorescence assay (IFA), and Western blot assays. The monoclonal antibodies will be used to develop validated diagnostic ELISA, IFA, Western blot tests and to better understand virus replication. Overall, the monoclonal antibodies, reagents, and assays will be vital in establishing the first early detection and surveillance tests thus preventing major economic losses to the swine industry.

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South Dakota State University


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