Staphylococcus aureus is one of the important pathogen often termed as superbug. After S. aureus interacting with host innate immune system, it induces the release of pro- inflammatory cytokines like TNF-α and IL-1β from Immune cells. In the current study, various strains of S. aureus and their cell wall lysates indicated cross reactivity with murine cytokines ELISA antibodies. In cell wall lysates of S. aureus, various proteins are involved in this cross reactivity phenomenon. This cross reactivity was exhibited in the form of unusual exaggerated signals while performing ELISA for quantification of murine cytokines. The proteins of cell wall lysates among various stains of S. aureus were determined using SDS-PAGE. To all of our data, it is revealed that unknown epitopes of S. aureus are involved in producing exaggerated signals of cytokines during their quantification by ELISA. This was generated not only during interaction of various strains of S. aureus with murine bone marrow derived macrophages (BMDM) but also in control settings where cell free system was used in ruling out this cross reactivity phenomenon.