Introduction: Urinary tract infection (UTI) is a common infectious disease which affects both, men and women. It is a significant health concern due to multidrug-resistant (MDR) organisms. Therefore, it is necessary to have a current understanding of the antibiotic susceptibility (AS) pattern of uropathogens to manage UTI effectively. Objective: This study aims to assess the antimicrobial sensitivity pattern of bacterial pathogens causing uncomplicated urinary tract infection in female patients. Methods: A prospective study was conducted at the Department of Pharmacology & Therapeutics in collaboration with the Department of Microbiology at SBMC, the Outpatient Department of Medicine, and Gynae & Obstetrics at Sher-E-Bangla Medical College, Barishal, Bangladesh, from January 2017 to December 2017. Clean catch midstream urine samples were collected and processed using standard guidelines for microbiological procedures. Positive microbiological cultures were found in 200 of the 314 patients, Data on socio-demographic, clinical, and risk factors were collected using a structured questionnaire. Results: In this study, the age of the subjects ranged from 15 to 75 years, majority of subjects (38.4%) belonged to age group of 45-60 years. The mean age was found 42.6±11.4 years. Out of 200 cases, E. coli was the most predominant gram-negative bacteria. Antibiotic susceptibility testing revealed that the pathogenic bacteria isolated from urine samples were MDR organisms. Aminoglycoside antibiotics such as Amikacin and gentamycin were the most effective drugs. Conclusion: Urinary tract infections are prevalent, affecting 64% of cases, with all isolates showing resistance to commonly used antibiotics. Therefore, it is recommended to provide health education on the transmission and causes of urinary tract infections.