Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions

Abstract

A 16S rRNA-based polymerase chain reaction (PCR) detection method was used to determine the prevalence of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Eikenella corrodens, Porphyromonas gingivalis, Prevotella intermedia. Prevotella nigrescens and Treponema denticola in subgingival specimens of 50 advanced periodontitis, 50 adult gingivitis and 50 pediatric gingivitis subjects. The optimal PCR conditions were determined for each study species. Agarose gel electrophoresis of PCR products from each study species revealed a single band of the predicted size. Restriction enzyme digestion of amplicons confirmed the specificity of the amplification. PCR detection limits were in the range of 25-100 cells. No cross-reactivity with other oral micro-organisms or nonspecific amplification was observed. The prevalence by PCR in advanced periodontitis, adult gingivitis and pediatric gingivitis subjects was 30%, 14% and 14% for A. actinomycetemcomitans, 86%, 18% and 8% for B. forsythus, 74%, 52% and 78% for C. rectus, 80%, 70% and 66% for E. corrodens, 70%, 10% and 14% for P. gingivalis, 58%, 12% and 18% for P. intermedia, 52%, 20% and 22% for P. nigrescens, and 54%, 16% and 16% for T. denticola, respectively. The prevalence was higher in the advanced periodontitis group than in both adult gingivitis and pediatric gingivitis for A. actinomycetemcomitans, B. forsythus, P. gingivalis, P. intermedia, P. nigrescens and T. denticola at P < 0.01, and for E. corrodens at P < 0.05. The prevalence of C. rectus was significantly higher in the advanced periodontitis group than in the adult gingivitis group at P < 0.01. Matching results between PCR and culture occurred in 28% (B. forsythus) to 71% (A. actinomycetemcomitans) of the samples; the major discrepancy occurred in the PCR-positive/culture-negative category. Matching results between PCR and DNA probe methods were found in 84% of the subjects (B. forsythus) and 70% (P. gingivalis). Odds ratio analysis revealed statistically significant positive associations between 17 of the 28 possible combinations (P < 0.01). This study demonstrated the utility of a 16S rRNA-based PCR detection method for identifying important subgingival microorganisms. The results indicated a strong association between the study species and periodontitis. Several previously unreported symbiotic relationships were found between the 8 species tested.

Biofilms of black tooth stains: PCR analysis reveals presence of Streptococcus mutans

“…The four analyzed bacteria (P. nigrescens, P. intermedia, S. mutans and Actinomyces spp) were detected in a similar prevalence in samples from patients with or without black tooth stains, suggesting a similar bacterial colonization of dental biofilms of these patients. According to the literature (5,20,21), among the 13 species of Prevotella, P. nigrescens and P. intermedia are the ones most frequently found in the oral cavity. P. nigrescens is indigenous microbiota of oral cavity whereas P. intermedia is related to periodontal disease.…”

PCR detection of four periodontopathogens from subgingival clinical samples

“…However, a suitable therapeutic to obtain a bacterial elimination may be of great clinical benefits (1). The use of the specific primers represent an appropriate tool to identify or detect periodontal pathogens from mixed bacterial populations.…”

Detection of putative periodontal pathogens in subgingival specimens of dogs

“…Moreover, it has been demonstrated a strong correlation between the presence of putative periodontal organisms and the destruction of periodontal tissues. Organisms such as Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Treponema denticola, Fusobacterium nucleatum, Dialister pneumosintes, Tannerella forsythensis, Treponema denticola, Eikenella corrodens and Campylobacter rectus have been related to several forms of periodontal disease in humans and animals (2,3,5,29).…”

“…Polymerase chain reaction (PCR) has been used in the direct identification of periodontal pathogens from subgingival specimens due to ability to accurately detect microbial species in mixed populations (3,4).…”