Methicillin-resistant Staphylococcus aureus is a clinically significant pathogen that causes infections ranging from skin and soft tissue infections to life-threatening sepsis. Biofilm formation by MRSA is one of the crucial virulence factor. Determination of beta-lactamase and biofilm production among Staphylococcus aureus was obtained from various clinical specimens. Standard bacteriological procedures were used for isolation and identification and antibiotic sensitivity was determined using the Kirby Bauer disc diffusion method according to CLSI guidelines. The cloverleaf method, acidometric, iodometric and chromogenic methods were used to detect beta-lactamase while the microtiter plate method and Congo red agar method were used to detect biofilm production. Of the 288 MRSA strains isolated from various clinical specimens,198 (67.07%) were biofilm producers. Cloverleaf and chromogenic (nitrocefin) disc shows 100% results for beta-lactamase detection. Vancomycin was 100% sensitive followed by teicoplanin (92.36%) and linezolid (89.93%). Cloverleaf and nitrocefin disc methods were the most sensitive for detection of beta-lactamase in S. aureus and there was no significant relation between biofilm production and antibiotic sensitivity pattern of S. aureus.