Infection by virulent strains of Mycobacterium tuberculosis (Mtb) induces a delayed type hypersensitive (DTH) reaction in all people who may, or may not develop the tuberculosis disease.  Immunization with Mycobacterium bovis, used in the BCG vaccine for TB, also produces the same DTH reaction.  Therefore, the tuberculin skin test, which measures the size of the skin induration following intradermal injection of tuberculin, cannot be applied on people who received prior BCG vaccination. We demonstrate here that JLEAPS-peptide heteroconjugates containing Mtb-specific peptides can be used in place of currently used ESAT-6 and CPF-10 proteins, in a restimulation assay with blood samples. We simultaneously assay a number of cytokines in order to reveal the distinction between Mtb- and BCG-exposed mice. Thus, the use of JLEAPS-peptide heteroconjugates containing Mtb-specific peptides in blood restimulation assay followed by simultaneous measurement of several cytokines may significantly enhance the sensitivity of Mtb detection in BCG-immunized subjects.