The invention relates to the technical field of auxiliary diagnosis of diseases, and particularly discloses an intraocular fluid cytokine detection reagent, a detection method and application. The detection reagent comprises at least two of the reagents for detecting Eotaxin, IL-10, IL-12p70, IL-1[beta], IL-2, IL-5, IL-6, IL-8, MCP-1, TNF, IFN-gamma, VEGF, IL-17a, ICAM-1, VCAM-1, TGF-[beta]1 and bFGF in the intraocular fluid. The detection method comprises the following steps: 1, uniformly mixing an intraocular fluid sample and a microsphere solution; 2, uniformly mixing the premixed sample and the antibody solution; 3, carrying out fluorescence detection on the to-be-detected sample through a flow cytometer. According to the application, the seventeen cell factors are screened and combined, so that the requirements of auxiliary diagnosis and typing of ophthalmic diseases can be met.