The present invention provides a hybridoma cell strain secreting monoclonal antibody against Tomato ringspot virus as well as antibody therefrom and antibody preparation method thereof, wherein: BALB/c mice are immunized with antigenic peptide CAFGKGVEEIEQTST to obtain a hybridoma cell strain, and the cell strain which can produce monoclonal antibody specifically recognize this antigenic peptide are screened, then monoclonal antibodies specifically recognizing Tomato ringspot virus coat protein AFGKGVEEIEQTST peptide are obtained by ascites preparation method, the lengths of heavy chain variable region and light chain variable region of this monoclonal antibody are respectively 185 and 103 amino acids； the yield of monoclonal antibody is 500-5000 mg/L ascites, the purity is 85%~99.5%. The monoclonal antibody of the present invention has a characteristic of specifically recognizing Tomato ringspot virus coat protein, and it has a broad application prospect in terms of Tomato ringspot virus entry-exit inspection and quarantine.
