Species-specific DNA probes were used to determine the presence of Actinobacillus actinomycetemcomitans (A.a.), Porphyromonas (Bacteroides) gingivalis, Prevotella intermedia, Treponema denticola, Eikenella corrodens, Fusobacterium nucleatum, and Wolinella recta in subgingival plaque from deep pockets/sites of patients with advanced periodontitis. The subjects were 20 patients with severe adult periodontitis, 13 men and 7 women (mean age 45.6 +/- 6.7 yr). For each subject, 9-10 subgingival sites with the deepest probing depths from each quadrant were sampled by the paper point method, a total of 198 sites, with mean probing depth 7.2 +/- 1.6 mm and clinical attachment level 9.5 +/- 2.7 mm. A.a. was present in at least one site in 75% of the subjects; P. gingivalis was found in 95%; P. intermedia and W. recta were found in 90%, respectively; and T. denticola, E. corrodens, and F. nucleatum were found in all subjects. In the 198 samples, A.a. was detected in 25.8%, P. gingivalis in 51.5%, P. intermedia in 64.1%, T. denticola in 60.6%, E. corrodens in 72.9%, F. nucleatum in 74.7%, and W. recta in 65.7%. The predominant combination was the simultaneous presence of P. intermedia, T. denticola, E. corrodens, F. nucleatum, and W. recta in 89.5% of the subjects and 46.8% of the sites. Of these sites, 51.1% showed the combined presence of P. gingivalis and 28.4% that of both A.a. and P. gingivalis. None of the seven bacteria could be detected in 14.4% of the total sites sampled. The present study indicates that severe destructive adult periodontitis is a multibacterial infection and that certain combinations of periodontopathogens seem to be important in the pathogenesis of the disease.