To evaluate procedures for the identification of Aerococcus urinae, we examined 24 alpha-hemolytic non-enterococcal bacterial isolates from 4373 urine samples. Published procedures were compared with 16s rRNA sequencing and biochemical profiling (BBL-Crystal-GP). 16s rRNA sequencing and BBL-Crystal-GP identified the same 13 isolates as A. urinae. Published tests failed to distinguish the 13 A. urinae isolates from eight non-A. urinae isolates; several tests exhibited no discrimination. Ciprofloxacin and trimethoprim susceptibility and growth at 45 degrees C improved discrimination. For urinary isolates, standard procedures for identification of A. urinae are redundant and insufficiently discriminatory, and may need revision. BBL-Crystal-GP is an accurate alternative.