Enterococci, which are commonly found in human fecal flora, have undergone a significant shift in recent years. Once viewed as minor clinical concerns for intestinal commensals, enterococci are now one of the most prevalent nosocomial pathogens, causing substantial morbidity and mortality. Aim: The objective of this study was to ascertain the emergence of multidrug-resistant enterococci isolated from a tertiary care teaching hospital in Vadodara. Materials & methods: In a retrospective analytical cross-sectional study conducted at a tertiary care center, 81 non-repetitive clinical isolates of enterococci were collected from a range of clinical samples and subjected to traditional phenotypic techniques, such as colony morphology, catalase test, bile esculin test ,automated Vitek 2 compact use to identify the isolates at the species level & to performed further antimicrobial susceptibility testing. The susceptibility and resistance of enterococci to antibiotics were determined based on recent the Clinical and Laboratory Standards Institute guidelines. Results: Enterococci isolates distributed by sample. 40 urine (49%) samples were collected, followed by blood (19%), then by pus (16%), and the others constituted 7%. study shows the sorting of enterococci isolates from different wards. In this study, 27% (n=22) of the patients from whom enterococci were isolated were from the NICU, 26% (n=21) from the MICU, and 47% (n=38) from various wards. Conclusion: Enterococcal isolates showed multidrug resistance, with E. faecium having a higher prevalence (53.08%) than E. faecalis (34.5%). Some isolates were resistant to all antibiotics tested, indicating that multidrug-resistant enterococci have become more common and pose a significant challenge for treatment. To address this issue, routine monitoring of antimicrobial resistance and implementation of effective infection control programs are essential. With a judicious antibiotic policy, the management of enterococcal infections can be improved.