The invention discloses an antigen composition used for detecting tuberculosis infection and application thereof. The antigen composition is composed of the first three or all of antigen epitope peptides of a CFP10 antigen, an ESAT6 antigen, a PPE antigen and an Ag85A antigen. The antigen composition is used for preparing a tuberculosis detection and diagnosis reagent and a kit used for detecting tuberculosis infection; the composition has the advantages of strong specificity and high sensitivity when being utilized for stimulating a specific T cell and detecting a chemokine secreted by the T cell; and the composition also has stronger specificity on diagnosis of tuberculosis infection, especially natural infection and latent infection such as bacillus of calmette and guerin vaccination.