Prulifloxacin, a fluoroquinolone antimicrobial agent which is a prodrug of ulifloxacin, has broad spectrum activity against gram positive and gram negative bacteria. In recent years it has been used efficaciously for the treatment of community acquired and nosocomial infections like urinary tract infections. The present study was aimed to compare the antimicrobial activity of prulifloxacin with other fluoroquinolones against urinary pathogens. The study was done of eighty urinary E. coli isolates. The result of this study indicates no significant difference in the antimicrobial activity was seen. The prolonged and high urinary concentration following a single oral dose strongly supports use of prulifloxacin for the treatment of urinary tract infections.