Staphylococcus epidermidis is considered as the main infectious agent associated with implanted medical devices. This study determined biofilm production and composition, pulsed-field gel electrophoresis (PFGE) profile, antimicrobial susceptibility and presence of virulence-related genes (ica operon, aap, bhp, embp, capB and IS256 transposase) in 49 clinical isolates of S. epidermidis. Twenty-five isolates (51%) were classified as biofilm producers in microtiter plate (MTP) assay. In Congo red assay (CRA) test, 14 (29%) showed positive reaction and three (6%) had indeterminate reaction, all were biofilmproducers in MTP assay. Fourteen isolates with positive reaction in CRA test had the chemical nature of biofilm determined as polysaccharide, had the ica operon (PIA-dependent producers) and the majority was strong biofilm producer. Eight biofilm producer isolates showed negative reaction in CRA test and the chemical nature of their biofilm was proteinaceous (PIA-independent producers). Antimicrobial resistance rates were generally higher in biofilm producers and resistance to beta-lactams ranged from 82-96%, while 61% of the isolates were multidrug resistant (≥ 10 drugs). Resistance to daptomycin, quinupristin/dalfopristin, rifampin and trimethoprim/sulfamethoxazole was observed only in PIAdependent isolates, while the resistance to gentamicin was present in all PIA-independent isolates and in just 53% of PIA-dependent of isolates. The most prevalent virulence-related genes were capB (80%) and embp (67%); the other genes were less frequent: ica operon (41%), aap (31%), IS256 transposase (22%) and bhp (10%). The presence of ica operon and IS256 transposase gene showed significant association with biofilm production and strong biofilm production. Moreover, these isolates presented significant higher resistance to levofloxacin, moxifloxacin, rifampin and trimethoprim/sulfamethoxazole. PFGE analysis showed 23 profiles, having the prevalent type 15 isolates. Of these, seven were PIAindependent biofilm producers and just one was PIA-dependent producer, unlike what was observed in other studies, where isolates of prevalent profiles were PIA-dependent biofilm producers.