The study of medicinal properties of plant based products is as old as medicine itself. Curative properties of herbal products have been exploited from the ancient past against various diseases. The recent trend of antibacterial research has revealed interest in search of new antibacterial agents of herbal origin that can have acceptability usually being nontoxic and inexpensive. The present study was undertaken to identify the strains of Staphylococcus aureus from the pus samples by using PCR technique. 16S rRNA gene was amplified using Universal primers. The amplified product was sequenced and the sequence was confirmed for Staphylococcus aureus after BLAST analysis with the NCBI database. Identified microorganisms were checked for their susceptibility to the Unani drug Itrifal Mundi. The microorganisms were found to be sensitive to the drug with the zone diameters ranging from 15mm to 22mm at the concentration of 10mg/ml. The MIC value of the drug was found to be in the range of 0.3125mg/ ml to 0.625mg/ml for most of the tested strains of S.aureus.