Urinary tract infections (UTIs) are among the most common maladies afflicting the human population globally. A wide variety of microbial pathogens are responsible for causing UTIs that are often recurrent, especially in the elderly population, resulting in an economic burden of billions of dollars annually. In this study, we present data from a large population of symptomatic UTI patients tested using a multiplexed, real time quantitative polymerase chain reaction (qRT-PCR) strategy. Our data demonstrate that instances of UTI increase with age for all the tested pathogens, with the exception of Mycoplasma spp. and Ureaplasma spp., which were found to be significantly higher in the younger population. Furthermore, the use of the qRT-PCR approach was also found to be effective in detecting polymicrobial UTIs. With the limitations of classical microbial culture techniques traditionally used for UTI diagnosis, we demonstrate that a rapid and comprehensive technique like RT-PCR can be an effective tool for detecting and managing UTIs.