BACKGROUND: In developing countries like Ethiopia, infections with antibiotic resistant bacteria become a real threat. Hence, monitoring of local level antimicrobial resistance profile is indispensable to contain the spread of drug resistant bacteria and intervene poor awareness on antimicrobial resistance. Therefore, this study aimed at determining bacterial and antibiotic resistance profiles of infections from different sites that occurred among patients. METHODS: Retrospective data recorded were analyzed on culture and drug susceptibility test results at Debre Markos Referral Hospital which were performed from 2011 to 2014. Drug susceptibility tests were performed using disk-diffusion technique. Chi square test was computed to compare the proportion of bacterial isolates with patients' age and sex. RESULTS: Out of 575 clinical samples processed, 280 (48.7%) were culture positive for aerobic bacteria pathogens. Wound 238 (41.4%) and urine 108 (18.8%) were the most frequent samples processed. Overall, Staphylococcus aureus (S. aureus) was the predominant isolate 100 (31.5%) followed by Escherichia coli (E. coli) 39 (13.8%), Pseudomonas aeruginosa (P. aeruginosa) 30 (10.3%) and Salmonella spp. 25 (8.9%). P. aeruginosa was the most frequent isolate followed by S. aureus from ear infection. E. coli was the leading isolate followed by Klebsiella spp. from urinary tract infection. Salmonella and Shigella spp. were the most frequent isolates in stool in children below 5 years of age. Neisseria gonorrhoeae (N. gonorrhoeae) 16 (76.2%) was the most common isolate from urethral discharge. The overall multidrug-resistant Gram positive and Gram negative bacteria isolates were 113 (84.6%) and 96 (72.2%), respectively. Gram positive bacteria revealed resistance to cotrimoxazole (80%), gentamicin (83.1%), amoxicillin (85.1%), ampicillin (85.8%), penicillin (89.7%), clindamycin (93.2%) and erythromycin (90.9%). Gram negative bacteria showed resistance to cotrimoxazole (53.1%), amoxicillin (58.8%), ampicillin (70.4%), tetracycline (75.9%) and gentamicin (76.9%). CONCLUSIONS: Various bacterial infections linked with high levels of MDR bacteria pathogens are major health problems in the study area. Therefore, treatment of common bacterial infections in the study area needs to be guided by drug-susceptibility testing of isolates.