Staphylococcus aureus (S. aureus) a noted versatile facultative commensal with pathogenic potential. This facultatively anaerobic cocci lives a commensal lifestyle in about a quarter of the world’s population awaiting a breach in the immune barrier to establish as a debilitating pathogen. This bacterium produces biofilms which acts as adhesive molecule enhancing the bacterial virulence and gives it an ability to withstand antibiotics, thereby causing tremendous burden on the healthcare community in terms of patient mortality and morbidity. This study assesses the biofilm producing ability among clinical isolates of staphylococci in comparison to colonizing staphylococci isolated from anterior nares of healthcare personnel. The present cross-sectional observational study was conducted in a tertiary care hospital in Mysore, India. In this study, 50 pathogenic isolates of S. aureus were assessed for phenotypic biofilm production. Additionally, 100 healthcare personnel were screened for nasal colonisation with S. aureus and the colonizing isolates were also screened for phenotypic biofilm formation. Phenotypic biofilm formation was evident among 22% of the pathogenic isolates. 23% of clinical S. aureus isolates demonstrated methicillin resistance (MRSA). There was no significant association of MRSA status with biofilm production (p > 0.05). Out of the 100 healthcare professionals screened for nasal carriage of S. aureus, 30% were colonized with S. aureus. Biofilm production among colonizing S. aureus isolates was found to be 26.6%. Methicillin resistance was a whopping 40% among the colonising strains. MRSA carriage was found to be highest among those healthcare personnel who had worked for an average of 20-30 years in the hospital environment. The Present study showed no significant association of biofilm production with the pathogenic potential of the S. aureus isolates (p > 0.05). Additionally, there was no significant association of methicillin resistance with the pathogenicity of the isolate. In conclusion, the interplay of virulence genes, biofilm forming ability, drug resistance of the isolate in coordination with the host immune status dictate the fate of the colonising as well as clinical isolates of S. aureus.