The invention discloses an incomplete antibody screening colloidal gold reagent kit and a preparation method thereof. The reagent kit comprises a first erythrocyte membrane antigen gold particle solution, a second erythrocyte membrane antigen gold particle solution, a third erythrocyte membrane antigen gold particle solution and an antihuman globulin solution, wherein an antigen spectrum of each of the first erythrocyte membrane antigen gold particle solution, the second erythrocyte membrane antigen gold particle solution and the third erythrocyte membrane antigen gold particle solution collectively include D, C, E, c, e, Jka, JKb, M, N, S, s, Fya, Fyb, Dia, K, k, Lea, Leb and P1 antigens. The reagent kit is long in storage time and easy to operate, the complicated process such as centrifuging, washing, and incubation is not needed, and a result is clear and easy to judge. The reagent kit also has the advantage of being suitable for large-scale operation and standard operation.