The invention discloses a test card for detecting glutamic-pyruvic transaminase by a photochemical method and a preparation method of the test card. The test card comprises a test card upper shell, atest card lower shell, a main test hole, a contrast test hole, a sample adding hole, a diffusion film, a first blood filtering film, two second blood filtering films, a first reaction film, a second reaction film and a single-sided adhesive tape, wherein the formula of a first reaction solution comprises L-alanine, pyruvate oxidase, alpha-ketoglutaric acid, peroxidase, thiamine pyrophosphate, magnesium chloride, 4-aminoantipyrine, a color developing agent, a film forming agent, a stabilizer and a buffer solution; the formula of a second reaction solution comprises pyruvate oxidase, peroxidase,thiamine pyrophosphate, magnesium chloride, 4-aminoantipyrine, a color developing agent, a film forming agent, a stabilizer and a buffer solution; by using the test card and the preparation method provided by the invention, endogenous pyruvic acid interference can be avoided, and the detection result is well matched with the hospital inspection result.