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Biochemical and genetic characterization of a Prevotella intermedia/nigrescens-like organism

¹Anaerobe Reference Laboratory, National Public Health Institute, Mannerheimintie 166, FIN-00300 Helsinki, Finland
²Institute of Dentistry, University of Helsinki, Helsinki, Finland
³Department of Oral Biology, Royal Dental College, University of Aarhus, Aarhus, Denmark
⁴VTT Biotechnology and Food Research, Espoo, Finland
⁵Wadsworth VA Medical Center, West Los Angeles, CA, USA

Author for correspondence: Eija Könönen. Tel: +358 9 4744 8248. Fax: +358 9 4744 8238.

ABSTRACT

Thirty-three previously non-typable faintly pigmented Gram-negative anaerobic bacterial isolates, biochemically most closely related to Prevotella intermedia and Prevotella nigrescens, were analysed for enzymic reactions, cellular fatty acid (CFA) composition, electrophoretic mobility of malate and glutamate dehydrogenases, hybridization with P. intermedia and P. nigrescens species-specific oligonucleotide probes and, for genetic heterogeneity, by arbitrarily primed PCR (AP-PCR). P. intermedia ATCC 25611^T and P. nigrescens ATCC 33563^T were run in parallel for comparison. Twenty-nine isolates originated from the normal oral flora of 18 subjects (including five mother-child pairs), and four isolates from various infections. Except for a negative lipase reaction, enzymic profiles of the test isolates were similar to those of P. intermedia and P. nigrescens. Clustering of CFAs, electrophoretic mobility patterns, hybridization with DNA probes for P. intermedia and P. nigrescens, and AP-PCR band patterns of the test isolates differed from those of the type strains of P. intermedia and P. nigrescens, suggesting the existence, in humans, of a new anaerobic species of pigmented, moderately saccharolytic, indole-positive Gram-negative rods.

Key Words: Prevotella intermedia/nigrescens-like organism • fatty acid profiles • arbitrarily primed PCR • multilocus enzyme electrophoresis • oral microorganisms

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