The invention discloses a CK, CKMB, LDH and AST combined detection reagent which comprises a diluent 1, a diluent 2 and 4 freeze-dried balls. The diluent 1 comprises trismetyl aminomethane, a surfactant, a dehydrobilirubin interference agent, vitamin C oxidase and a preservative; the diluent 2 comprises a buffer solution ad an inhibitory CK-M antibody; the freeze-dried ball 1 comprises a buffer solution, alpha-ketoglutaric acid, a reduced coenzyme I, malic dehydrogenase, L-aspartate and a freeze-drying protective additive; the freeze-dried ball 2 comprises a buffer solution, L-lithium lactate,an oxidized coenzyme I and a freeze-drying protective additive; the freeze-dried balls 3 and 4 comprise a buffer solution, adenosine diphosphate, glucose-6-phosphate dehydrogenase, hexokinase, D-glucose, phosphocreatine, an enzyme activator and a freeze-drying protective additive. The combined detection reagent is applicable to a multifunctional full-spectrum POCT (Point-of-care Testing) biochemical analyzer, has good correlation to detection results of clinically common liquid reagents on a large biochemical analyzer, and has the advantages of being simple and convenient to operate, convenient in reagent preservation and transport, low in detection cost and the like.