The new Vitek 2 GN card (bioMérieux, Marcy-l'Etoile, France) was developed for better identification of fermenting and nonfermenting bacilli. This new card allows the identification of 159 taxa. A total of 426 isolates (331 fermenting and 95 nonfermenting gram-negative bacilli) belonging to 70 taxa covered by the database were evaluated. All isolates were identified in parallel with the ID 32 GN, the API 20E, and the API 20NE methods. The system correctly identified 97.4% (n=415) of the strains. Only 2.1% (n=9) needed additional testing. One strain (0.25%) was misidentified (Klebsiella pneumoniae subsp. pneumoniae), and another one (0.25%) was not identified (Morganella morganii subsp. morganii). The new GN card gives more accurate identifications overall for gram-negative bacilli when compared to the systems described in other similar studies.