The diagnosis and follow up of patients with bladder cancer relies heavily on the use of invasive procedures. Hyaluronic acid and hyaluronidase may serve as markers for bladder cancer since they are involved in the biology of it. Accordingly, urinary HA (ELISA) and HAase RNA (RT-PCR) were measured in the present study to examine the usefulness of these markers. In our comparative study (n=268 cases, 162 malignant), RT-PCR for urinary hyaluronidase RNA had superior sensitivity (91.4%), specificity (93.4%), accuracy (92.2%) than urinary HA (ELISA assay) 78.4%, 70.2%, 75.6% respectively, or urine cytology (67.3%, 80.2%, 72% respectively) in detecting bladder cancer. Overall the urinary hyaluronidase RNA appears to be a promising method of detecting new onset of bladder tumor and evaluating its grade than others.