The invention discloses an envelope protein VP28 idiotype monoclonal antibody against shrimp white spot syndrome virus (WSSV) and a preparation method thereof. The antibody is secreted by a hybridoma cells with the collection number of CCTCC-CT200938, is prepared by taking anti-WSSV-VP28 monoclonal antibody (Ab1) as antigen, can bind with anti-WSSV-VP28 antibody of hare, and has the capability of competing with WSSV to bind with the anti-WSSV-VP28 antibody of the hare. The anti-WSSV-VP28 idiotype monoclonal antibody (Ab3) prepared by taking the antibody as antigen can bind with the WSSV, the binding site of the anti-WSSV-VP28 idiotype monoclonal antibody (Ab3) and the WSSV is located on an envelope, and the Ab3 can neutralize WSSV infection and has Ab1 properties. In the invention, the idiotype antibody is applied in the research of WSSV for the first time; a screening system is established, which uses an indirect enzyme-linked immunosorbent assay (ELISA) method and a competitive enzyme-linked immunosorbent assay (ELISA) method for detection; the fact that Ab3 has properties of Ab1 is proved by adopting an indirect immnnofluotesent method (IIF), a gold labeling immunoelectron microscopic method and crayfish in vivo neutralization tests, thus proving that the monoclonal antibody in the invention has the property to simulate original antigen WSSV-VP28.