The invention discloses an antigenic polypeptide pool capable of detecting mycobacterium tuberculosis infection. The antigenic polypeptide pool specifically stimulates mycobacterium tuberculosis infected fresh whole blood to specifically secrete IFN-gama and increase the sensitivity. The invention provides a new detection reagent for detecting mycobacterium tuberculosis infection, experiments prove that peripheral blood can be directly utilized to conduct antigenic simulation without separation of peripheral blood mononuclear cell, the experimental data shows that the antigenic polypeptide pool has high sensitivity and specificity for detecting mycobacterium tuberculosis infection, and is simple and convenient to operate and low in cost, thus having high clinical application values.