To determine the pattern of microbes responsible for urinary tract infections and their susceptibility to different antibiotics. MethodThis is a cross-sectional study conducted at Quetta, Pakistan. The urine samples of 400 patients were collected and sent for culture and sensitivity analysis. The results were recorded on an excel datasheet. Descriptive statistics were used to describe the data. ResultsOut of 400 urine samples, 266 samples were culture positive for microorganisms. The most common organism on analysis was Escherichia coli 123/266 (46.24%) followed by Staphylococcus saprophyticus 59/266 (22.18%) and Klebsiella pneumonia 49/266 (18.42%). Gram-negative microorganisms were most susceptible to fosfomycin, cefoperazone/sulbactam, and meropenem. Gram-positive microorganisms were most susceptible to fosfomycin, cefoperazone/sulbactam, meropenem, and amoxicillin/clavulanate. High rates of resistance in E. coli were observed to most commonly prescribed broad-spectrum antibiotics; ceftriaxone (64.35%), cefotaxime (76.54%), ceftazidime (49.43%), cefepime (53.44%), levofloxacin (71.26%), and amoxicillin/clavulanate (70.31%). E. coli was the major multidrug-resistant organism. ConclusionHigh rates of antibiotic resistance and multi-drug resistance were revealed in this study due to the widespread and injudicious use of broad-spectrum antibiotics. Thus, it is highly recommended to regulate the pharmacies. Physicians should judiciously prescribe antibiotics and practice the culture and sensitivity of urine samples rather than blind prescription. Continued surveillance on uropathogens prevalence and resistance, new and next-generation antibiotics, and rapid diagnostic tests to differentiate viral from bacterial infections is the need of time.