VITEK 2 is an automated identification system for diverse bacterial and fungal species. A new card (the Neisseria-Haemophilus [NH] card) for the identification of Neisseria spp., Haemophilus spp., and other fastidious gram-negative or gram-variable microorganisms has been developed, but its performance in a routine clinical laboratory has not yet been evaluated. In this study, a total of 188 bacterial strains belonging to the genera Actinobacillus, Campylobacter, Capnocytophaga, Cardiobacterium, Eikenella, Gardnerella, Haemophilus, Kingella, Moraxella, and Neisseria were investigated. The NH card was able to identify 171 strains (91%) correctly without the need for extra tests; one strain (0.5%) was misidentified, and five strains (2.7%) could not be classified. Eleven strains (5.8%) were identified with a low level of discrimination, and simple additional tests were required to increase the correct-identification rate to 96.8%. The results were available within 6 h. Based on these results, the new VITEK 2 NH card appears to be a good method for the identification of diverse groups of fastidious organisms, which would otherwise require testing with multiple systems. However, more work is needed to evaluate the performance of VITEK 2 with regard to Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella bacteria because of the insufficient number of strains tested in this study. Moreover, further reduction of the detection time would be desirable.Fastidious bacteria are frequently associated with severe infectious diseases. In particular, Neisseria and Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella (HACEK) organisms are responsible for such serious infectious conditions as sepsis, meningitis, and endocarditis (5,7,11,13,14,17,22,23,26). These organisms are sometimes difficult to cultivate and identify. The VITEK 2 system (bioMérieux, Marcy L'Etoile, France) is used worldwide for the rapid and accurate identification of gram-positive cocci, gram-negative rods, and yeasts (1,2,16,18,24) in routine clinical microbiology tests. BioMérieux has recently developed a new Neisseria-Haemophilus (NH) identification card in order to identify fastidious gram-negative and gram-variable organisms (HACEK organisms), but its performance under the conditions of a routine laboratory has not yet been evaluated. The new VITEK 2 NH system allows for the identification of 28 taxa belonging to the genera Actinobacillus, Campylobacter, Capnocytophaga, Cardiobacterium, Eikenella, Gardnerella, Haemophilus, Kingella, Moraxella, Neisseria, Oligella, and Suttonella. The NH card is based on colorimetric technology and uses dehydrated media containing chromogenic substrates, which are detected by optical heads. A computer-assisted algorithm based on data collected from known strains of the claimed species is used to interpret the data for a final identification. In this study, 188 isolates belonging to the species included in the database of the VITEK 2 system's software were used to evaluate the ability of ...