The utility of the 16S ribosomal RNA-based polymerase chain reaction (PCR) for detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Eikenella corrodens, Porphyromonas gingivalis, Prevotella intermedia, and Treponema denticola was examined and compared with that of anaerobic culture. Primer pairs consisting of 20-27 nucleotides amplified 404- to 688-bp regions of 16S ribosomal RNA genes of these organisms. This method had a lower detection limit of 50 target cells in a background of 10(7) cells. Its specificity for B. forsythus, P. gingivalis, and T. denticola seemed high. The primers for A. actinomycetemcomitans, C. rectus, and P. intermedia cross-reacted with some closely related species but did not reveal amplification products in tests with more distantly related organisms. The primers for E. corrodens did not seem to cross-react with oral organisms. This PCR technique was sensitive, reproducible, and easy to perform. PCR-based amplification may prove valuable for the detection of some periodontal pathogens in crude subgingival specimens.

Anaerobic bacteria play important roles in the pathogenesis of human periodontitis. This study examined the relationship between a potentially new periodontopathic bacterium Dialister pneumosintes and periodontal disease. A total of 73 women and 62 men aged 18 to 86 years participated in the study. Using a 16S rRNA polymerase chain reaction identification method, the presence of D. pneumosintes was determined in paper-point samples from periodontal pockets of 105 periodontitis and 30 gingivitis patients. D. pneumosintes was detected in 83% of patients having severe periodontitis and in 19% of patients having slight periodontitis. We suggest adding D. pneumosintes to the group of suspected periodontal pathogens.

An open-label, multicenter study was performed to assess bacteriologic findings associated with chronic bacterial maxillary sinusitis in adults. Seventy aerobic (52.2%) and 64 anaerobic (47.8%) pathogens were recovered from clinically evaluable patients at baseline (before therapy). The most commonly isolated anaerobes were Prevotella species (31.1%), anaerobic streptococci (21.9%), and Fusobacterium species (15.6%). The aerobes most frequently recovered included Streptococcus species (21.4%), Haemophilus influenzae (15.7%), Pseudomonas aeruginosa (15.7%), and Staphylococcus aureus and Moraxella catarrhalis (10.0% each). Recurrences of signs or symptoms of bacterial maxillary sinusitis associated with anaerobes were twice as frequent as were those associated with aerobes when counts of anaerobes were > or =10(3) cfu/mL. A pathogenic role for Granulicatella species in cases of chronic sinusitis was documented for the first time.

This study examined the effect of oral food consumption on the prevalence and levels of subgingival bacteria and yeasts in 20 gastrostomy tube‐fed children and 24 healthy controls. Microbial identification was carried out using anaerobic culture and 16S rRNA‐based PCR identification methods. Streptococcal and Actinomyces species were recovered from 100% and 76% of all subjects and averaged 66% and 11% of total cultivable organisms, respectively. In decreasing order of prevalence, Fusobacterium, enteric rods, Prevotella intermedia/Prevotella nigrescens, Capnocytophaga, Propionibacterium, yeasts, Actinobacillus actinomycetemcomitans, coagulase‐negative Staphylococcus, Campylobacter rectus, Bacteroides forsythus, and Porphyromonas gingivalis were detected in 48% to 2% of the study subjects. The cultivable levels of these species varied widely among subjects. PCR detection showed C. rectus and Eikenella corrodens both to occur in 93% of the study subjects and to be the most prevalent putative periodontal pathogens examined. In decreasing order of prevalence, PCR identified Treponema denticola, A. actinomycetemcomitans, P. nigrescens, P. intermedia, B. forsythus, and P. gingivalis in 38% to 21% of the subjects studied. Tube‐fed children and healthy controls exhibited similar subgingival microbial compositions. It appears from this study that oral food consumption is not a major determinant for the establishment of subgingival microbiota in children. J Periodontol 1997;68:1163–1168.

This study compared the ability of a nonradioactive digoxigenin-labeled DNA probe and anaerobic culture to identify subgingival Porphyromonas gingivalis. Total cellular DNA from P. gingivalis ATCC 33277T was labeled using the Genius kit from Boehringer Mannheim Biochemicals. Anaerobic culture was performed using VMGA III transport medium and enriched brucella blood agar. The DNA probe could detect as little as 1000 P. gingivalis cells added to supragingival plaque. Also, the probe could detect P. gingivalis when it was present in proportions too low to be visualized on overgrown bacterial plates. The probe showed no visible reaction with strains of various oral species or with thousands of non-P. gingivalis colonies from plaque samples. VMGA III could maintain the viability of P. gingivalis for up to 6 days, as evidenced by DNA probing of colony blot of subgingival cultures. A total cellular DNA probe for detecting P. gingivalis seems to offer a simple and reliable method of detecting the organism in subgingival specimens.