A rapid DNA probe method for detection of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans

BACKGROUND: The objective of the present study was to develop a rapid DNA probe method for the microbiological detection of periodontitis that can be used in dental clinics. By using the DNA probe, we also investigated the correlation between the occurrence of putative periodontopathic bacteria and clinical parameters. METHODS: This rapid DNA probe method minimizes the use of a water bath for ordinary hybridization and washing in order to shorten the total reaction time. The detection process could be completed within 2 hours. In order to evaluate the clinical application of the DNA probe, subgingival plaque samples were taken from patients with periodontitis before initial therapy. After the therapy, the patients were microbiologically and clinically evaluated. RESULTS: When the DNA probe method was compared with the culture method, the agreement was 88% for Porphyromonas gingivalis and 67% for Actinobacillus actinomycetemcomitans. A statistically significant association was found between the detection of P. gingivalis and probing depth, bleeding on probing (chi2 test: P <0.001, P <0.05). A significant association was also shown between the detection of A. actinomycetemcomitans and probing depth in patients aged 35 or older (chi2 test: P <0.001). The detection rate of A. actinomycetemcomitans was highest in teenagers. At shallow periodontal pocket sites (PD < or =3 mm) in teenagers, no P. gingivalis was found, while 22% of the sites harbored A. actinomycetemcomitans. After the therapy, the frequency of detection of P. gingivalis decreased significantly only in the clinically improved sites (chi2 test: P <0.001). CONCLUSIONS: The rapid DNA probe method appears promising as an efficient tool for rapid clinical detection of periodontopathic bacteria.     

Identification of periodontopathic bacteria DNA from periodontitis-involved gingival tissue using a PCR method

The purpose of this study was to determine the presence of periodontopathic bacteria DNA in the granulation tissue of periodontally involved gingival tissue. Forty periodontitis patients were examined. Subgingival plaque and saliva samples were collected from the patients before surgery. Then, granulation tissue was also collected during surgery. Another 20 patients, who had more than 4 mm in probing pocket depth, were also examined as a control. They received only initial treatment without periodontal surgery. Plaque and saliva samples were also collected from them. Seven periodontopathic bacteria, Porphyromonas gingivalis, Bacteroides forsythus, Toreponema denticola, Campylobacter rectus, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Prevotella nigrescens, were detected by a PCR method from these samples of all patients. As clinical parameters, probing pocket depth, clinical attachment level, and bleeding on probing were recorded. One year after surgery, sampling of subgingival plaque and saliva, and measurement of clinical parameters were performed on the patients. The DNA of periodontopathic bacteria was detected in the granulation tissue of 29 out of 40 patients. P. gingivalis, B. forsythus, T. denticola, C. rectus, A. actinomycetemcomitans, P. intermedia and P. nigrescens were detected in 23, 11, 12, 9, 3, 2 and 1 subject(s) respectively. The sites with undetectable level of any periodontal pathogens at one year after surgery showed a clinical attachment gain of 2.44 mm, while those that showed detectable pathogen levels had only a 0.77 mm gain. Our results indicated that the removal of granulation tissue contributed to eliminating periodontal pathogens and to avoiding re-infection by those bacteria. Complete eradication of periodontopathic bacteria would be necessary for successful periodontal treatment.     

Microbiological markers for prediction and assessment of treatment outcome following non-surgical periodontal therapy

BACKGROUND: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Bacteroides forsythus are considered major putative periodontal pathogens. However, it remains unclear what combinations or what levels of these bacteria influence treatment outcome. The purpose of the present study was to establish useful pathogenic markers for prediction and assessment of treatment outcome following scaling and root planing (SRP). METHODS: A total of 1,149 sites in 104 chronic periodontitis patients were clinically examined at baseline. Three months after SRP, 606 sites in 56 of these patients were reexamined. Subgingival plaque samples taken from the examined sites at baseline and 3 months were analyzed for the detection and quantification of A. actinomycetemcomitans, P. gingivalis, and B. forsythus using a colorimetric polymerase chain reaction technique. RESULTS: At baseline, high levels and a combination of P. gingivalis and B. forsythus were frequently detected in diseased sites (74%). SRP reduced the levels and the coexistence of P. gingivalis and B. forsythus (from 75% to 43%). However, in treated sites where there was less reduction of probing depth (<2 mm), or where bleeding on probing (BOP) or suppuration was detected, residual coexistence of P. gingivalis and B. forsythus and a high level of P. gingivalis after SRP were significantly more frequent. Furthermore, SRP did not improve BOP at sites exhibiting initially high levels of A. actinomycetemcomitans. CONCLUSIONS: These results suggest that the combination of P. gingivalis and B. forsythus, as well as the level of P. gingivalis, is useful in assessing treatment outcome. Furthermore, the high level of A. actinomycetemcomitans before SRP is a possible valuable predictor of treatment outcome.     

Porphyromonas gingivalis,Bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction

BACKGROUND AND AIMS: Bacteria play an essential role in the pathogenesis of destructive periodontal disease. It has been suggested that not all bacteria associated with periodontitis may be normal inhabitants of a periodontally healthy dentition. In particular, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans have been isolated infrequently from subjects without periodontitis. The aim of the present study was to compare prevalence and proportions of a number of periodontal bacteria in periodontitis patients and control subjects. MATERIAL AND METHODS: In all, 116 consecutive subjects diagnosed with moderate to severe periodontitis (mean age 42.4) and 94 subjects without radiographic evidence of alveolar bone loss (mean age 40.4) were recruited for the study. The gingival condition in the control group varied between gingival health and various degrees of gingivitis. In patients, the deepest pocket in each quadrant was selected for microbiological sampling. In control subjects all mesial and distal sites of all first molars were selected for sampling. All paper points from a patient were pooled and processed for anaerobic cultivation within 6 h after sampling. Clinical variables of sampled sites included bleeding index, probing pocket depth and clinical attachment level. RESULTS: A. actinomycetemcomitans, P. gingivalis, Prevotella intermedia, Bacteroides forsythus, Fusobacterium nucleatum and Peptostreptococcus micros were significantly more often prevalent in patients than in controls. The highest odds ratios were found for P. gingivalis and B. forsythus (12.3 and 10.4 resp.). Other odds ratios varied from 3.1 to 7.7 for A. actinomycetemcomitans and P. micros, respectively. Absolute numbers of target bacteria were all higher in patients, but only the mean percentage of B. forsythus was significantly higher in patients in comparison to controls (P < 0.001). CONCLUSIONS:A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus, F. nucleatum and P. micros are all significant markers for destructive periodontal disease in adult subjects. Based on calculated odds ratios, B. forsythus and P. gingivalis are the strongest bacterial markers for this disease and are infrequently cultured from subjects without periodontal bone loss.     

Transmission of periodontal disease-associated bacteria from teeth to osseointegrated implant regions

PURPOSE: The presence of periodontopathic bacteria is a risk factor for peri-implantitis. The present study examined colonization by periodontopathic bacteria and their transmission from periodontal pockets to osseointegrated implant sulcus. MATERIALS AND METHODS: Plaque samples were collected from 105 sites in the 15 patients who participated in the study. Colonization by these bacteria was examined by polymerase chain reaction (PCR) and culture. The transmission of periodontopathic bacteria from periodontal sites of natural teeth to the implant sulcus was analyzed by pulsed field gel electrophoresis (PFGE). RESULTS: The PCR detection rates of Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Treponema denticola were 80.0%, 53.3%, 46.7%, 60.0% and 40.0%, respectively. Colonizations by P gingivalis and A actinomycetemcomitans were statistically correlated with periodontal pockets and implant sulcus regions (P < .01). The PFGE patterns of the P gingivalis strains isolated from each patient were identical, but differed from those from other patients. The PFGE patterns of P intermedia strains were identical in 2 out of 3 patients. DISCUSSION: These analyses indicated that there appeared to be transmission of P gingivalis and P intermedia from the periodontal pocket to the peri-implant region. CONCLUSION: Elimination of these periodontal pathogens from the patient's oral cavity before administering dental implant treatment may inhibit colonization by these pathogens and reduce the risk of peri-implantitis.     

Microbiological response of localized sites with recurrent periodontitis in maintenance patients treated with tetracycline fibers.

BACKGROUND: Whether adjunctive tetracycline fibers can provide an additive effect to scaling and root planing in treating non-responsive sites in maintenance subjects is still controversial. Recolonization of the bacteria from untreated sites or from the extracrevicular region may explain the insignificant response to local therapy. The purpose of the present study was to evaluate the microbiological response of sites treated with tetracycline fibers combined with scaling and root planing. METHODS: The study was conducted in a split-mouth design. Thirty patients on maintenance therapy having at least 2 non-adjacent sites in separate quadrants with probing depths between 4 to 8 mm with bleeding on probing, or aspartate aminotransferase enzyme levels > 800 microIU in the gingival crevicular fluid, were treated with scaling and root planing plus tetracycline fibers or with scaling and root planing only. Subgingival plaque samples were collected at baseline, and 1, 3, and 6 months following treatment. A. actino-mycetemcomitans, C. rectus, B. forsythus, E. corrodens, F. nucleatum, P. gingivalis, and P. intermedia were detected by culture, immunofluorescence, or PCR technique. RESULTS: There was a reduction of total bacterial cell count, as well as of certain periodontal pathogens, following treatment. The prevalence of A. actinomycetemcomitans, B. forsythus, and P. gingivalis and the mean proportions of C. rectus, P. intermedia, F. nucleatum, and P. gingivalis decreased after therapy, but there was no statistically significant difference between the 2 treatment groups with respect to bacterial proportions or the number of positive sites. Besides, the pathogens could not be eliminated from the periodontal pocket, and recolonization of the pocket was noted at 3 months post-treatment. CONCLUSIONS: Bacteria located within the cheek, tongue mucosa, saliva, or untreated sites may contribute to reinfection of the pockets and explain the insignificant response to local tetracycline therapy.     

Relationship between herpesviruses and adult periodontitis and periodontopathic bacteria.

BACKGROUND: Various mammalian viruses and specific bacteria seem to play important roles in the pathogenesis of human periodontitis. This study examined the relationship between subgingival herpesviruses and periodontal disease and potential periodontopathic bacteria in 140 adults exhibiting either periodontitis or gingivitis. METHODS: A nested-polymerase chain reaction (PCR) method determined the presence of Epstein-Barr virus type 1 and type 2 (EBV-1, EBV-2), human cytomegalovirus (HCMV), and herpes simplex virus (HSV) and a 16S rRNA PCR detection method identified Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola. RESULTS: Using a logistic analysis, EBV-1 showed significant positive association with P. gingivalis (odds ratio [OR] 3.37), and with coinfections of P. gingivalis and P. intermedia (OR 4.03); P. gingivalis and B. forsythus (OR 3.84); P. gingivalis and T. denticola (OR 4.17); P. gingivalis, B. forsythus, and T. denticola (OR 4.06); and P. gingivalis, P. nigrescens, and T. denticola (OR 3.29). EBV-1 also showed positive association with severe periodontitis (OR 5.09), with increasing age (OR 1.03), and with periodontal probing depth at the sample sites (OR 1.77). HCMV was positively associated with coinfections of P. gingivalis and P. nigrescens (OR 3.23); P. gingivalis, B. forsythus, and P. nigrescens (OR 3.23); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.59); with severe periodontitis (OR 4.65); and with age (OR 1.03). Patients with mixed viral infections revealed significant associations with P. gingivalis (OR 2.27), and with coinfections of P. gingivalis and B. forsythus (OR 2.06); P. gingivalis and P. nigrescens (OR 2.91); P. gingivalis, B. forsythus, and P. nigrescens (OR 2.91); and P. gingivalis, P. nigrescens, and T. denticola (OR 2.70) with the clinical diagnosis of slight (OR 3.73), moderate (OR 3.82), or severe periodontitis (OR 4.36), and with probing depth at the sample sites (OR 1.39). HSV and EBV-2 showed no significant associations with any of the variables tested. CONCLUSIONS: The results indicate that subgingival EBV-1, HCMV, and viral coinfections are associated with the subgingival presence of some periodontal pathogens and periodontitis. Herpesviruses may exert periodontopathic potential by decreasing the host resistance against subgingival colonization and multiplication of periodontal pathogens.     

Detection of periodontopathic bacteria and an oxidative stress marker in saliva from periodontitis patients

We assessed the salivary levels of periodontopathic bacteria and 8-hydroxydeoxyguanosine (8-OHdG) in patients with periodontitis. The salivary levels of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia (formerly Bacteroides forsythus) were assessed using real-time polymerase chain reaction. The 8-OHdG levels were determined using an enzyme-linked immunosorbent assay. The salivary levels of 8-OHdG, P. gingivalis, and T. forsythia in the periodontitis patients were significantly higher than those in healthy subjects. By contrast, the A. actinomycetemcomitans level in healthy subjects was higher than that in periodontitis patients. 8-OHdG was significantly correlated with P. gingivalis. Statistically significant decreases in the levels of P. gingivalis, probing depth, bleeding on probing, and 8-OHdG were observed after initial periodontal treatment. These results suggest that the 8-OHdG levels in saliva reflect the load of periodontal pathogens. 8-OHdG could be a useful biomarker for assessing periodontal status accurately, and for evaluating the efficacy of periodontal treatment.     

Comparative analysis of putative periodontopathic bacteria by multiplex polymerase chain reaction

Background and Objective:  The polymerase chain reaction (PCR) has been applied for the rapid and specific detection of periodontopathic bacteria in subgingival plaque and is potentially of clinical benefit in the diagnosis and treatment of periodontitis subjects. However, several technical points need to be modified before the conventional PCR detection system can be used by clinicians.
Material and Methods:  To develop a PCR-based technique more applicable for clinical use than conventional PCR, we established a multiplex PCR for five putative periodontopathic ( Treponema denticola , Porphyromonas gingivalis , Aggregatibacter actinomycetemcomitans , Prevotella intermedia and Tannerella forsythia ) and two nonperiodontopathic ( Streptococcus sanguinis and Streptococcus salivarius ) species of bacteria using whole-plaque suspension as templates, and detected bacteria in subgingival plaque taken from 85 subjects at the supportive periodontal therapy stage after active periodontal treatments.
Results:  Among putative periodontopathic bacteria, the detection frequency of T. denticola and P. gingivalis was elevated in parallel with higher probing pocket depth and clinical attachment loss, and had 4.2–14.1 times increasing odds of the clinical parameters tested. Detection of any of the five species of putative periodontopathic bacteria markedly increased the odds ratio of a higher probing pocket depth, clinical attachment loss and bleeding on probing.
Conclusion:  The multiplex PCR system developed in this study enabled the detection of all the bacteria under investigation in one reaction tube in a less time- and labor-intensive manner than conventional PCR. These results support the potential clinical use of multiplex PCR for detecting periodontopathic bacteria and for evaluating therapeutic strategies and predicting the prognosis for each subject.     

Sulfate-reducing bacteria in relation with other potential periodontal pathogens

BACKGROUND, AIMS: Oral sulfate-reducing bacteria are involved in several clinical categories of periodontitis. The aim of this cross-sectional study was to compare the presence of sulfate-reducing bacteria (SRB) with other putative pathogens including spirochetes, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, and Treponema denticola in periodontal lesions. METHOD: Periodontal SRB were detected by enrichment culture and compared with a microscopic spirochete count (n=168). Species-specific oligonucleotide probes directed against the 16S rRNA were employed to determine the presence of A. actinomycetemcomitans, P. gingivalis, B. forsythus, and T. denticola (n=55). RESULTS: A significant positive correlation was observed between the presence of SRB and the proportions of spirochetes in subgingival plaque, although the 2 bacterial groups also occurred separately. SRB tended to be negatively correlated with the presence of A. actinomycetemcomitans. In contrast, all pockets with SRB harbored either T. denticola, or both T. denticola and B. forsythus (12/14) before therapy. Interestingly, the combination of SRB with P. gingivalis occurred in 32% of the periodontal pockets before treatment. After initial periodontal therapy, the prevalence of this combination was reduced to 2% of the sites, and to 25% of the sites in recall patients. CONCLUSION: The presence of SRB was positively correlated with T. denticola, B. forsythus, and P. gingivalis in periodontal lesions. These suspected pathogens form a complex strongly associated with destructive periodontitis.     

Occurrence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in progressive adult periodontitis

BACKGROUND: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia are the major periodontal bacteria species in most forms of progressive periodontitis in Scandinavia and the United States. The occurrence of periodontal pathogens appears to be different in subjects of different ethnic origin, and geographical factors may influence the distribution of these species. METHODS: The occurrence of A. actinomycetemcomitans, P. gingivalis, and P. intermedia was determined using a DNA probe in progressive adult periodontitis in Chileans. Sixty patients (mean age 43.6 +/- 8 years) who had not previously received any type of periodontal therapy were selected. Bleeding on probing, probing depth, and clinical attachment level measurements were made with an automated probe. Patients were monitored at 2-month intervals until at least 2 sites exhibited > or =2 mm attachment loss. Two subgingival plaque samples from active sites were taken in 56 subjects and matched with 2 plaque samples from inactive sites in the same individuals. RESULTS: P. gingivalis was found in 75% of active sites and in 59.7% of inactive sites in 96% of the patients (P = 0.022). P. gingivalis at high levels of detection was significantly more frequent in active sites (48.2%) than in inactive sites (31.2%) (P = 0.014). A. actinomycetemcomitans was detected in 6.25% of active sites and in 12.5% of inactive sites in 11.6% of patients. P. intermedia was found in 33% of patients and at a significantly higher proportion in active sites (49.1%) than in inactive sites (30.3%) (P = 0.006). There was a significantly higher proportion of inactive sites (34.8%) than active sites (19.6%) without any of the 3 pathogens (P = 0.016). Bleeding on probing was significantly more associated with active sites with high levels of P. gingivalis and with active sites with P. intermedia than with inactive sites. CONCLUSIONS: A high prevalence of P. gingivalis and P. intermedia was found in adult periodontitis, and the occurrence of these bacteria appears to be higher in Chileans than in other populations. No apparent association exists between A. actinomycetemcomitans and progressive adult periodontitis in Chileans.     

Clinical and microbiologic changes after removal of orthodontic appliances.

The goal of this study was to evaluate the clinical and microbiological factors associated with orthodontic appliances during an episode of gingival inflammation and the impact of appliance removal on periodontal health. This prospective study included 10 patients, aged 12 to 20 years, with clinical signs of gingival inflammation at the final phase of orthodontic treatment (appliance removal). Plaque index, gingival index, and probing depth were evaluated, and microbiological samples were collected from teeth 16, 11, and 26 at 2 times: during the gingival inflammation (baseline) and 30 days after the removal of the appliance and professional prophylaxis. Polymerase chain reaction analysis was used to detect Porphyromonas gingivalis, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Prevotella nigrescens. A statistically significant improvement of the plaque and gingival indexes was seen, as well as a reduction in probing depth (P <.05). Periodontal pathogens were associated with gingival inflammation during orthodontic treatment. The improvement in periodontal health at 30 days was concomitant with a reduction of sites positive for A. actinomycetemcomitans and B. forsythus (P <.05). Periodontal pathogens associated with gingival inflammation during orthodontic treatment can be significantly reduced by orthodontic appliance removal and professional prophylaxis.     

Subgingival distribution of periodontal pathogenic microorganisms in adult periodontitis.

The association between specific plaque microorganisms and periodontal diseases has been the subject of much recent interest due to its potential importance in the diagnosis and classification of these diseases. In order to optimize microbiological tests in periodontal therapy, it is important to know how many subgingival plaque samples must be assayed from a single patient in order to ascertain infection with a periodontal pathogen. To answer this question the present study assessed the distribution of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, and Prevotella intermedia in multiple subgingival plaque samples. The samples were quantitatively assessed for specific bacteria by indirect immunofluorescence, a technique previously found to correlate well with cultural assessment of these same organisms. Subgingival plaque from the mesial pockets/sulci of all teeth except third molars was sampled in 12 patients with adult periodontitis, 22 to 28 sites/patient for a total of 315 samples. These patients demonstrated an average mesial probing depth and loss of attachment of 3.7 +/- 1.2 mm and 3.1 +/- 1.5 mm, respectively (mean +/- SD). P. gingivalis, P. intermedia, and B. forsythus were demonstrated in one or more sites from all patients, while A. actinomycetemcomitans was found in one or more sites in 8 of 12 patients. The proportion of positive sites per subject varied, but it was, on average, similar for the 3 black-pigmented organisms and ranged from 44% to 54%. In contrast, A. actinomycetemcomitans was identified, on average, in only 11.4% of the sites in these same patients.(ABSTRACT TRUNCATED AT 250 WORDS)     

Initial effect of controlled release chlorhexidine on subgingival microorganisms

BACKGROUND: Little or no data exist on the ability of subgingival application of PerioChip (2.5 mg chlorhexidine gluconate in a biodegradable chip; Astra Pharmaceuticals, Westborough, MA, USA) to suppress periodontopathic microorganisms. The present study compared the subgingival microbiota of periodontitis sites receiving the chlorhexidine chip plus scaling and root planing (Sc/Rp) or Sc/Rp alone. METHODS: Seven males and six females, mean age 49 years, with moderate to advanced periodontitis participated in the study. In each patient, two bilateral pockets probing 6-7 mm were randomly assigned to treatment by chlorhexidine chip + Sc/Rp, or by Sc/Rp alone. Subgingival placement of chlorhexidine chips was carried out according to the manufacturer's instructions. Sc/Rp was performed with hand instruments for at least 10 min in each study tooth. Subgingival samples were collected by paper-points at baseline, at 2 weeks and at 4 weeks post-treatment. Anaerobic culture methods were used for microbial isolation and identification. The microbiologic examination was carried out blindly. Microbiological data were evaluated by a repeated measures analysis of variance. RESULTS: No statistical difference was found in total colony counts between subgingival sites treated with chlorhexidine chip + Sc/Rp and those treated with Sc/Rp alone. Also, the percentage of major periodontal pathogens (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Bacteroides forsythus) and the percentage of total periodontal pathogens (A. actinomycetemcomitans, P. gingivalis, B. forsythus, Prevotella intermedia-group, Fusobacterium, Eubacterium, Campylobacter rectus, Peptostreptococcus micros, Eikenella corrodens, enteric rods) were not significantly different between the chlorhexidine chip + Sc/Rp group and the Sc/Rp group. At baseline, A. actinomycetemcomitans was recovered from 4 chlorhexidine chip + Sc/Rp sites and 2 Sc/Rp sites, P. gingivalis from 5 chlorhexidine chip + Sc/Rp sites and 4 Sc/Rp sites, and B. forsythus from 9 chlorhexidine chip + Sc/Rp and 7 Sc/Rp sites. At 4 weeks, A. actinomycetemcomitans was detected in 2 chlorhexidine chip + Sc/Rp sites but not in any site receiving Sc/Rp, P. gingivalis in 2 chlorhexidine chip + Sc/Rp sites but not in any Sc/Rp site, and B. forsythus in 1 chlorhexidine chip + Sc/Rp and in 2 Sc/Rp sites. CONCLUSION: The present data obtained from bilateral periodontitis lesions of 13 adults suggest that chlorhexidine chip treatment of adult periodontitis lesions provides little or no additional antimicrobial benefits compared to thorough Sc/Rp alone.     

Periodontopathic bacterial infection in childhood

BACKGROUND: Little information is available on periodontopathic bacterial infection in childhood. We assessed the prevalence by age of 10 putative periodontopathic microorganisms in periodontally healthy children using a polymerase chain reaction (PCR) assay. METHODS: Plaque samples were collected from the buccal-mesial sulcus of the first molar or second primary molar in the right upper quadrant of 144 children (2 to 13 years old, 12 subjects from each year of age) who showed negligible periodontal inflammation. Using species-specific primers of Porphyromonas gingivalis, Bacteroides forsythus, Prevotella intermedia, Prevotella nigrescens, Campylobacter rectus, Eikenella corrodens, Actinobacillus actinomycetemcomitans, Capnocytophaga ochracea, Capnocytophaga sputigena, and Treponema denticola, PCR amplification was performed with bacterial genomic DNA from plaque samples. RESULTS: The results indicated that C. rectus, E. corrodens, A. actinomycetemcomitans, C. ochracea, and C. sputigena were found in about 50% of the plaque samples from all age groups, while B. forsythus and P. intermedia were detected less frequently, and P. gingivalis and T. denticola were not found. In contrast, the percentage of P. nigrescens-positive subjects increased with age in primary dentition, and reached about 50% at 7 years of age and older. Subject-based analyses suggested that the number of bacterial species in the plaque samples increased gradually with age until 5 years old, and then reached a plateau after the mixed dentition period. CONCLUSIONS: The colonization of many putative periodontopathic microorganisms can occur quite early in childhood without clinical signs of periodontal disease. However, colonization by P. gingivalis and T. denticola was not detected in periodontally healthy children.     

Detection of Tannerella forsythensis (Bacteroides forsythus) and porphyromonas gingivalis by polymerase chain reaction in subjects with different periodontal status

BACKGROUND: The aim of this study was to determine the prevalence of Tannerella forsythensis (formerly Bacteroides forsythus) and Porphyromonas gingivalis in subgingival plaque samples by using polymerase chain reaction (PCR), and to assess the relationship of these bacteria with different categories of periodontal disease and health. METHODS: Subjects were distributed into 3 groups according to their periodontal diagnosis: group 1, periodontally healthy (N = 10); group 2, periodontitis with probing depth < or = 5 mm (N = 10); group 3, periodontitis with probing depth > 5 mm (N = 10). The subjects in groups 2 and 3 had healthy and diseased periodontal sites. Subgingival plaque samples were obtained using paper points inserted into periodontal pockets (diseased sites) and into healthy gingival sulci (healthy sites) of the same subject. RESULTS: The distribution of bacteria differed in healthy and diseased sites. T. forsythensis (B. forsythus) was not detected in any sample from healthy sites in any group but was detected in 70% and 100% of diseased sites in groups 2 and 3, respectively. P. gingivalis was detected in only one sample from a healthy site (group 2), and in the diseased sites, its prevalence was 40% (group 2) and 90% (group 3). In addition, T. forsythensis (B. forsythus) and P. gingivalis were both detected in 30% and 90% of the diseased sites in groups 2 and 3, respectively. CONCLUSION: These results indicate a possible association between periodontal disease and the presence of T. forsythensis (B. forsythus) and/or P. gingivalis.     

Prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in subgingival microflora of Japanese patients with adult and rapidly progressive periodontitis

BACKGROUND, AIMS: This study investigated the prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans among various periodontitis patients and healthy individuals in Japan, and correlated it with clinical parameters. METHOD: Subgingival plaque samples were collected from 21 patients with adult periodontitis (AP), 8 with rapidly progressive periodontitis (RPP) and 15 healthy individuals. RESULTS: The frequency detected in culture was as follows: B. forsythus was found in 47.6% of AP sites and in 37.5% of RPP sites. P. gingivalis was identified in 64.3% of AP and 59.4% of RPP sites. A. actinomycetemcomitans was detected in 4.8% of AP and 3.1% of RPP sites. The 3 species were detected in only 2 of the healthy individuals. The proportion of B. forsythus in the total microflora in culture was 0.07% in the healthy group, 4.1% in AP and 2.4% in RPP. The proportions of P. gingivalis were 0% in the healthy group, 18.8% in AP and 16.2% in RPP. The proportion of A. actinomycetemcomitans was very low in all 3 groups. A DNA probe detected B. forsythus in 78.6% of AP and 65.6% of RPP sites, as well as P.gingivalis in 58.3% of AP and 59.4% of RPP sites. A. actinomycetemcomitans was detected in only 1.2% of AP sites. The 3 species were undetectable in the healthy group. CONCLUSIONS: The prevalence and the proportion of B. forsythus and P. gingivalis were significantly correlated with clinical parameters, suggesting that B. forsythus and P. gingivalis are closely related to AP and RPP in the Japanese population.     

Periodontopathic bacteria in children with Down syndrome

BACKGROUND: It is widely known that individuals with Down syndrome (DS) often develop severe early-onset periodontal diseases. In this study, we examined the prevalence of periodontopathic bacteria in DS children to determine if specific pathogens are acquired in their childhood. METHODS: The subjects were 60 DS children (2 to 13 years old, 5 in each age bracket) and 60 age-matched controls. Ten pathogens, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, Treponema denticola, Prevotella intermedia, P nigrescens, Capnocytophaga ochracea, C. sputigena, Campyrobacter rectus, and Eikenella corrodens were surveyed in subgingival plaque samples using a polymerase chain reaction. Periodontal status was evaluated by probing depth, bleeding on probing, and gingival index. RESULTS: No significant difference in periodontal status was observed between the DS and control groups, however, all of the pathogens were detected with greater frequency in the DS children. B. forsythus, T. denticola, P. nigrescens, and C. rectus were significantly prevalent throughout all age brackets of the DS children (P <0.01 or 0.05). The occurrence of P. gingivalis was also significant in the DS subjects over 5 years old. A cluster analysis of the microbial profiles of the DS subjects showed that gingivitis severity was associated with increased varieties of the harboring pathogens and the distribution of P. gingivalis. CONCLUSIONS: These results suggest that various periodontopathogens can colonize in the very early childhood of DS patients and maturation of subgingival components, including P. gingivalis, plays an important role in the initiation of gingival inflammation.     

Prevalence of periodontopathic bacteria in aggressive periodontitis patients in a Japanese population

BACKGROUND: Actinobacillus actinomycetemcomitans is considered a major etiologic agent of aggressive periodontitis. Other periodontopathic bacteria such as Porphyromonas gingivalis are also suspected of participating in aggressive periodontitis, although the evidence is controversial. The aim of the present study was to investigate the prevalence of periodontopathic bacteria and to clarify the microbiological features of aggressive periodontitis in Japanese patients. METHODS: Subgingival plaque was collected from 50 aggressive periodontitis (AgP) patients (localized 10, generalized 40). Samples from 35 generalized chronic periodontitis (CP) patients and 18 healthy subjects were examined as controls. Plaque samples were examined using culture and polymerase chain reaction (PCR) method. RESULTS: The prevalence of A. actinomycetemcomitans was relatively low in the localized (20%) and generalized (17.5%) AgP patients, with no significant difference observed in detection frequencies between AgP and the control groups (CP 8.6%, healthy 0%). On the other hand, Tannerella forsythensis (formerly Bacteroides forsythus), Campylobacter rectus, P. gingivalis, and Treponema denticola were frequently detected in localized as well as generalized aggressive periodontitis patients. The prevalence and proportion of P. gingivalis correlated with severity of clinical attachment loss in both localized and generalized aggressive periodontitis. CONCLUSIONS: T. forsythensis, C. rectus, P. gingivalis, and T. denticola were the predominant periodontopathic bacteria of aggressive periodontitis patients in Japan. Although A. actinomycetem- comitans was also detected in AgP patients, the prevalence of this bacterium was much lower than that of P. gingivalis.