Urinary tract infection (UTI) is one of the most important causes of morbidity in the general population. A total of 2400 midstream urine samples were processed from patients having clinically suspected UTIs attending various hospitals in Pattukkottai from January 2012 to June 2013. Uropathogens were identified by conventional methods. Antimicrobial susceptibility was done by the disc diffusion methods. The commercially available 16 different types of antimicrobial discs were used. Over all incidences of UTIs in the present study was higher in females (69.8%) than in males (30.2%). The most common isolates in this study were Gram negative bacilli which accounts for 85.4% of the total positive isolates. Major Gram negative isolates were E.coli (54.6%); K. pneumoniae (11.2%) was the second major isolates and Pseudomonas spp. (10.5%) was the third major isolates. In Gram positive isolates, Coagulase-negative Staphylococci, Enterococcus and Staphylococcus saprophyticus were the least dominant uropathogens causing UTI strains. This study showed that antibiotics like imipenem, amikacin, nitrofurantoin and gentamicin are still very effective. While an increased resistance to amoxycillin, ampicillin, cotrimoxazole and norfloxacin were observed. A total of twenty two numbers of Candida species were isolated, of which C. albicans showed the highest number of isolates. Amphotericin B were the most effective antifungal agent to all Candida isolates. The multi-drug resistant ESBL producing E.coli isolates were present in these area hospitals with a prevalence of 10%. There were two predominant genes TEM and SHV detected among MDR E.coli uropathogenic strains circulating in these area hospitals. E.coli was found one of the commonest causative agents of UTIs. The higher percentage of incidence of E.coli showed the significant health threat to the patients in the study area. Hence, the present findings will be useful for the clinician to provide safe and effective empirical antibiotic therapies to treat UTIs.