The invention discloses an antigen polypeptide pool for detecting Mycobacterium tuberculosis infection. The specific antigen polypeptide pool can specifically stimulate Mycobacterium-tuberculosis-infected fresh whole blood and specifically secrete IFN-gamma, thereby increasing the detection sensitivity. The invention provides a novel detection reagent applicable to detection of Mycobacterium tuberculosis infection. The experiment proves that the peripheral blood can be directly utilized to perform antigenic stimulation without the need for separating peripheral blood mononuclear cells. The experimental data shows that when being used for detecting Mycobacterium tuberculosis infection, the antigen polypeptide pool has the advantages of higher sensitivity, higher specificity and lower cost, is simple to operate, and has higher clinical application value.