Our study was aimed to generate data on prevalence of antibiotic resistance among clinically significant bacteria isolated from patients at a tertiary care hospital in Chandrapur, Maharashtra, India. Various clinical samples were processed for culture, identification and AST by conventional method. Among 265 different isolates, 54.3% were Gram positive and 45.6% were Gram negative. The commonest pathogen isolated was Staphylococcus aureus, followed by E.coli. Among S. aureus, 51.9% were MRSA. Most of Gram positive bacteria were resistant to co–trimaxozole and ciprofloxacin while most Gram negative bacilli were resistant to ampicillin, amoxyclav, co–trimoxazole and cephalosporins. Least resistance was for amikacin. The study showed a high rate of MRSA infection along with in general resistance amongst gram positive and negative isolates to commonly used drugs underlining need of continuous surveillance, antimicrobial stewardship and strict infection control program to restrict the ongoing resistance.