Background Enterococci are common bacteria found in the human mouth, stomach, and female vaginal system that are known to cause nosocomial infections. Urinary tract infections, followed by intra-abdominal abscesses and bloodstream infections, are the most common illnesses caused by Enterococcus spp.Material methodThis prospective observational study was conducted in department of microbiology. Clinical samples such as urine, pus, cerebrospinal fluid and other body fluid were received in department of microbiology, mahatma Gandhi memorial medical college, Indore Madhya Pradesh from February 2019 to January 2020. In this duration 200 Enterococcus species isolates were isolated. The antimicrobial susceptibility testing was done by Kirby Bauer’s discdiffusion method as per 2018 CLSI guideline.Result In this study, 31453 clinical samples received in the bacteriology laboratory in which 2832 morphological suspected colonies were subjected to biochemical testing, 200 isolated strains were found Catalase Negative, Bile Esculin positive, Growth in 6.5% NaCl, Growth at 10℃ and 45℃ and sugar fermentation. Species of Enterococci, E. faecalis constituted 155/200 (77.5%), and E. faecium 45/200 (22.5%).Conclusion The finding of this study indicated the presence of Enterococcus species have shown an increased rate of resistance to most the of tested drug, particularly to Vancomycin. This finding demands an attention from health policy makers for intensified actions to promote rational use of antibiotics in health care settings and surveillance studies in order to monitor changes in Enterococcal resistance patterns.