The invention discloses a kit for determination of alpha-
hydroxybutyrate dehydrogenase and a preparation method thereof, wherein the kit includes double-liquid components of a
reagent R1 and a
reagent R2 which are independent with each other and includes the components and the corresponding content: the
reagent R1 including 1-350 mmol / L of a buffer liquid, 0.01-10 mmol / L of alpha-
ketone butyric acid, 0.1-2 g / L of disodium
ethylenediamine tetraacetate, 0.01-0.40 g / L of a stabilizing agent, and a
solvent being
purified water; and the reagent R2 including 1-350 mmol / L of a buffer liquid, 0.1-4.0 mmol / L of NADH, 0.01-0.40 g / L of a stabilizer, and a
solvent being
purified water. The preparation method includes the steps: according to the following component content, preparing the reagents; mixing a to-be-tested sample with the reagent R1 and the reagent R2, and carrying out full reaction; determining the
absorbance difference value after the reaction with a
fully automatic biochemical analyzer; and calculating the concentration of the alpha-
hydroxybutyrate dehydrogenase in the sample according to the
absorbance change value. The kit has the advantages of high accuracy, good precision and the like.