Present study reports the development of simple, user friendly qualitative cum quantitative biosensor based on urease enzyme for the analysis of urea. Enzyme was isolated from a urease producing microbe originally isolated from National Fertilizer Ltd., Bathinda, which was later found to be a Corynebacterium spp. (MTCC 8143), having log phase between 9-24 hrs. Highest enzyme activity was found to be at 21 hrs. To fabricate biosensor crude enzyme extract and indicator (phenol red) was co-immobilized on nylon membranes with TEOS based hydrosol-gel. Membranes with immobilized biocomponent and indicator when dipped in urea solution, color changed from yellow to red-violet. Time taken for the change in color (response time) was correlated with urea concentration. Urea could be detected in the solution with a detection limit as low as 10-9M with a response time 2 min 29 seconds. Constructed biosensor was applied to human blood and urine samples. Bio-component was stable for 20 days when kept at 40C.