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Nonhemolytic variants ofHaemophilus haemolyticus are difficult to differentiate fromHaemophilus influenzae despite a wide difference in pathogenic potential. A previous investigation characterized a challenging set of 60 clinical strains using multiple PCRs for marker genes and described strains that could not be unequivocally identified as either species. We have analyzed the same set of strains by multilocus sequence analysis (MLSA) and near-full-length 16S rRNA gene sequencing. MLSA unambiguously allocated all study strains to either of the two species, while identification by 16S rRNA sequence was inconclusive for three strains. Notably, the two methods yielded conflicting identifications for two strains. Most of the “fuzzy species” strains were identified asH. influenzae that had undergone complete deletion of the fucose operon. Such strains, which are untypeable by theH. influenzae multilocus sequence type (MLST) scheme, have sporadically been reported and predominantly belong to a single branch ofH. influenzae MLSA phylogenetic group II. We also found evidence of interspecies recombination betweenH. influenzae andH. haemolyticus within the 16S rRNA genes. Establishing an accurate method for rapid and inexpensive identification ofH. influenzae is important for disease surveillance and treatment.

journal of clinical microbiology

Complete Deletion of the Fucose Operon in Haemophilus influenzae Is Associated with a Cluster in Multilocus Sequence Analysis-Based Phylogenetic Group II Related to Haemophilus haemolyticus: Implications for Identification and Typing