Mouse monoclonal antibodies specifically recognizing the Penicillin Binding Protein 2a (PBP2a) derived from a strain of Methicillin-Resistant Staphylococcus aureus (MRSA) were produced and characterized. The immunogen used to generate an immune response in a mouse was a PBP2a recombinant protein derived from a strain of Methicillin-Resistant Staphylococcus aureus (MRSA). The data showed that both monoclonal antibodies of the disclosure were able to distinguish MRSA from MSSA bacteria. The monoclonal antibodies have distinct recognition patterns for the regions of the PBP2a protein sequence. Epitope mapping has localized regions of the PBP2a protein specifically recognized by one or both of the monoclonal antibodies. The monoclonal antibodies of the present disclosure having the ability to distinguish between MRSA and MSSA strains can be useful as the basis for a diagnostic assay useful in the clinical setting for determining whether and which antibiotics to administer to a patient.