Identifying clinically important gram-negative anaerobes from the oral cavity

Gram-negative oral anaerobes have frequently been associated with periodontal disease, some species more frequently than others. The confusing classification of these organisms has often obscured the association with disease of particular species within this group of organisms. This investigation aimed to compare different identification methods that could be applied in clinical research. Clinical isolates were collected and identified by three different methods: screening with phenotypic tests, commercial identification kits, and a 16S rRNA-based polymerase chain reaction (PCR) method. Forty-three reference strains of 19 Prevotella and Porphyromonas species were also included in the investigation. The phenotypic screen easily differentiated Porph. gingivalis from the other pigmented species. The screen also gave a good indication of separation of the lactose-fermenting species from the lactose non-fermenting species, although diversity can be seen in beta-galactosidase activity. Commercial identification kits did not add much to identification achieved with the phenotypic screen, only 20% of Porph. gingivalis isolates could be identified to species level with the kits. Neither the kits nor the phenotypic screen could differentiate Pr. intermedia and Pr. nigrescens. With the PCR method, Pr. intermedia and Pr. nigrescens were easily separated, and Porph. gingivalis was readily identified. Because of 16S rRNA gene sequence similarity, Pr. melaninogenica and Pr. veroralis could not be separated by the PCR method.