MDR uropathogens lead to high-level morbidity and mortality and pose a tremendous threat towards treatment failure and survival of UTI patients. This study was conducted to evaluate the demographic characteristics, antibiogram, and resistance pattern of Escherichia coli against antibiotics-a retrospective study at Nishtar Medical Hospital in Multan, from June 2016 to July 2017. A total of 150 clinical samples were collected and biochemically characterized by Berge's manual, and antimicrobial susceptibility testing was performed by using the Kirby-Bauer disc diffusion method and the results were interpreted according to Clinical and Laboratory Standards Institute standards. Data was analyzed by the XL-stat software. Results indicate that UTI was more prevalent in rural areas such as Alipur (28.6%) and Jalalpur (18%) and less common in urban areas like Muzaffargarh (2.66) and Multan (2%). A total of 150 patients were examined, of which 88 were females and 62 males in which more frequently found pathogens were E. coli (33%) and others Klebsiella pneumonia (18%), Proteus (10%), Staphylococcus saprophyticus (8.66%), Staphylococcus aureus (7.33%), Pseudomonas aeruginosa (6.66%), Enterobacter (6%), Candida (5.33%), and Citrobacter (4.66%). The antimicrobial susceptibility testing was carried out against E. coli strains that showed the highest overall resistance patterns of imipenem (IMI), Cip, Aug, Cfm, and CN, which were 80%, 72%, 68%, 60%, and 52%, respectively. MDR E. coli strains showed the highest resistance pattern towards IMI which put through alarming situation in Pakistan. Significant results showed that TZP, AK, and F were found sensitive in vitro against E. coli isolates of UTIs. Prevalence of MDR uropathogens near around Multan has been first time documented in this study with respect to possible risk factors associated with UTI. These alarming signs of MDR E. coli have been rarely addressed and warrant attention on urgent basis.