The invention relates to the technical field of enzyme linked immune, and discloses a composition for an enzyme linked immunosorbent assay kit, an anti-nuclear antibody spectrum detection kit and a preparation method thereof. The composition comprises sealing liquid and ELISA diluents; the sealing liquid contains casein, saccharose, PBS, mannitol, Tween20, lycine and sodium azide; the ELISA diluents contain PBS, casein, sodium p-hydroxybenzoate, PEG, lycine and Proclin300. By starting from the sealing liquid and the ELISA diluents of the enzyme linked immunosorbent assay kit, the proper ingredients are selected, so that the enzyme linked immunosorbent assay kit can maintain the detection stability for a long time. Meanwhile, the anti-nuclear antibody spectrum detection kit prepared from the composition has higher stability; the guarantee period is 2 years or more.