Microbial comparison of smoker and non-smoker adult and early-onset periodontitis patients by polymerase chain reaction

A number of bacterial species are involved in the aetiology of periodontitis and include Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus and Treponema denticola. Several studies have shown differences in the microflora between the various forms of periodontal disease. It is recognised that smoking is a risk factor for periodontal disease, but there are conflicting reports on whether or not smoking has an effect on the periodontal microflora. We utilised the polymerase chain reaction to determine the presence of A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus and T. denticola in subgingival plaque samples in 33 adult periodontitis (AP) patients and 24 generalized early-onset periodontitis (GEOP) patients prior to treatment. When GEOP and AP patients were compared there were significant differences in the number of positive patients and sites for both A. actinomycetemcomitans and B. forsythus (p=0.0023 and 0.00001, respectively). No statistically significant differences in the prevalence of these organisms were found between smoker and non-smoker groups. These results confirm that AP and GEOP sites harbour varied microflora, but show that B. forsythus and A. actinomycetemcomitans were detected to a significantly greater extent in this group of GEOP than in the AP patients investigated. Our findings do not support the hypothesis that smokers have significant differences in the prevalence of periodontal pathogens from non-smokers.     

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.     

Periodontal pathogens in the shallow pockets of immigrants from developing countries

The aim of this study was to examine the distribution of typical periodontitis-associated microorganisms in refugees arriving from non-industrialized countries, and to relate the presence of these organisms to the periodontal condition of the subjects. Thirty males between 35-44 years of age were surveyed. Dental plaque, calculus, gingivitis, loss of attachment, and probing depths were recorded for all surfaces. A total of 90 microbiological samples were taken with paper points from mesial sites of teeth 16, 36 and 41. Microbiological test sites were grouped by probing depths and loss of attachment. Only 16.8% of all surfaces had probing depths > 3 mm, although 90.7% of surfaces had loss of attachment > 1 mm. Twenty-one sites with obvious periodontal destruction (PD > 3 mm, LA > 2 mm) showed the greatest recovery of Porphyromonas gingivalis (66.7%). However, 51 sites with minimal periodontal disease (PD < or = 3 mm, LA < 2 mm) and with no gingival recession also showed a relatively high detection frequency of P. gingivalis (34.1%). Twenty-four of these samples came from 12 patients with no pockets > 5 mm and with less than 10% of all sites yielding pockets > 3 mm. The detection frequencies of Prevotella intermedia (91.6%), Bacteroides forsythus (25.0%), Wolinella spp. (33.3%) and Actinobacillus actinomycetemcomitans (50.0%) were similar in these sites compared with periodontitis sites. Morphologically distinct isolates, from 19 individuals positive for A. actinomycetemcomitans, were serotyped by indirect immunofluorescence.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevalence of Bacteroides forsythus and Bacteroides gingivalis in subgingival plaque of prosthodontically treated patients on short recall

The prevalence of Bacteroides forsythus and Bacteroides gingivalis in subgingival plaque of patients on short recall was analyzed in relation to the probing depth of the test sites. The subjects had excellent oral hygiene and therefore were unlikely to suffer from active periodontal destruction. Sixty-four subgingival plaque samples, taken from gingival or periodontal pockets with probing depths ranging from 1 to 8 mm, were quantitatively assessed for the presence of the two species using species-specific monoclonal antibodies in conjunction with a very sensitive indirect immunofluorescence technique. Both organisms were encountered in probes from sites as shallow as 2 mm, but the percentage of positive samples clearly rose in relation to the probing depth of the test sites. Overall, B. forsythus was found to colonize lesions earlier, that is at smaller probing depths, than B. gingivalis. Interestingly, whenever a sample was found to be positive for B. gingivalis it was also positive for B. forsythus. The numbers of B. forsythus and B. gingivalis and the total bacterial cell number found in the pockets were significantly correlated to the probing depth. However, with advancing probing depth the increase of the total cell numbers of the two Bacteroides species was considerably more pronounced than the increase of the total subgingival plaque cell number. The recall interval neither affected the frequency of sites positive for B. forsythus or B. gingivalis nor influenced significantly the proportions of the two species in subgingival plaque.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of major putative periodontopathogens in Korean advanced adult periodontitis patients using a nucleic acid-based approach

BACKGROUND: Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systematic analysis of subgingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to describe the prevalence of major putative periodontopathogens in Korean patients by culture-independent methods. METHODS: A total of 244 subgingival plaque samples (5 sites in each participant) were taken from 29 advanced adult periodontitis (AP) patients and 20 periodontally healthy subjects. AP samples were obtained from the 4 deepest periodontal pockets (> or =6 mm probing depth [PD]) and 1 healthy site (< or =3 mm PD) in each patient. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed with eubacterial primers. Aliquots of PCR products were then applied on nylon membranes and hybridized with specific oligonucleotide probes labeled with digoxigenin. RESULTS: All diseased sites harbored Fusobacterium sp., while Porphyromonas gingivalis, Treponema sp., and Bacteroides forsythus were detected in more than 96% of 116 diseased sites. Peptostreptococcus micros, Actinobacillus actinomycetemcomitans, and Prevotella intermedia were present in 82%, 74%, and 71% of diseased sites, respectively. In sites of periodontally healthy subjects, Fusobacterium sp. was present in the highest proportion (58%). Treponema sp., P. gingivalis, and B. forsythus were detected in 22%, 18%, and 18% of healthy sites, respectively. P. micros, P. intermedia, and A. actinomycetemcomitans were found in 8%, 2%, and 1% of healthy sites, respectively. The prevalence of the periodontopathogens, with the exceptions of Fusobacterium sp. and B. forsythus, was significantly higher in the healthy sites of periodontitis subjects than in the healthy sites of periodontally healthy subjects (P <0.05). CONCLUSIONS: Using highly sensitive methods relying on 16S ribosomal RNA-based oligonucleotide probes, we confirmed the strong association of 7 putative periodontopathogens with AP patients in a Korean population. With the exceptions of Fusobacterium sp. and B. forsythus, all the periodontopathogens were significantly more associated with the healthy sites of periodontitis subjects than in the healthy sites of periodontally healthy subjects.     

Predominant microflora of severe, moderate and minimal periodontal lesions in young adults with rapidly progressive periodontitis

The purpose of this investigation was to study the microflora of severe, moderate and minimal periodontal lesions, in young adults with rapidly progressive periodontitis (RPP). Subgingival plaque samples were taken from 142 periodontal lesions in 10 young adults aging 25 to 35 years. The examination of the subgingival microflora indicated that certain species, including Porphyromonas gingivalis, Bacteroides forsythus, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans , and Campylobacter species were found to be predominant in severe periodontal lesions. B. forsythus, P. gingivalis, Prevotella intermedia, F. nucleatum, Capnocytophaga ochracea , were predominant in medium lesions while Streptococcus species and Actinomyces species, C. ochracea, Haemophilus segnis and Veillonella parvula , were found in higher levels in minimal periodontal lesions.     

Multi-center clinical evaluation of a chairside method for detecting certain periodontopathic bacteria in periodontal disease

The association of bacteroides gingivalis, Bacteroides forsythus, Treponema denticola, and Actinobacillus actinomycetemcomitans among others with periodontal disease offers the opportunity for the development of diagnostic tests that are based upon the detection and/or quantification of one or more of these organisms or their by-products in the plaque. Three of the putative periodontal pathogens namely, T. denticola, B. gingivalis, and B. forsythus, can hydrolyze the synthetic trypsin substrate, N-benzoyl-DL-arginine-2-naphthylamide (BANA) forming a color reaction. The present investigation evaluated a commercially developed solid state assay for BANA hydrolysis that can be read after 15 minutes incubation at chairside. A total of 702 subgingival plaque samples were collected from 117 patients seen at four university dental clinics and placed on reagent cards. The color development on the cards was compared to the presence of T. denticola and B. gingivalis in the plaque, and with the clinical appearance of the sampled sites. This multi-center study demonstrated that antibodies to B. gingivalis and T. denticola could detect these organisms by an ELISA in the majority of the subgingival plaque samples. Comparable information could be obtained when the same plaques were evaluated by the reagent card format for BANA hydrolysis. The ELISA and reagent card were comparable in their ability to distinguish between clinically healthy and diseased sites. Both diagnostic procedures detected the periodontopathogens in plaques from sites that were judged clinically healthy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Persistence patterns of Porphyromonas gingivalis, Prevotella intermedia/nigrescens, and Actinobacillus actinomyetemcomitans after mechanical therapy of periodontal disease

BACKGROUND: The aim of this study was to determine the distribution patterns of Porphyromonas gingivalis, Prevotella intermedia/nigrescens, and Actinobacillus actinomycetemcomitans in periodontitis patients after standard mechanical periodontal therapy, and to determine factors increasing the odds to detect these target organisms in treated sites. METHODS: Eight hundred fifty-two (852) separate subgingival microbial samples were taken from the mesial and distal aspects of every tooth in 17 patients. Target organisms were identified culturally. RESULTS: The 3 microorganisms showed different persistence patterns: P. gingivalis was detected in a high percentage of subjects (59%), but in a low proportion of sites (5.4%). P. intermedia/nigrescens was detected in all subjects except one, and in 40.6% of the tested sites. Only 5 subjects were A. actinomycetemcomitans positive, but 2 of them showed a very high number of positive sites (44% and 75%, respectively). A highly significant relationship was found between a subject's tendency to bleed upon sampling and the number of P. intermedia/nigrescens-positive sites. A significant portion of the variation in frequency of persisting P. gingivalis could be explained by the frequency of persisting pockets deeper than 4 mm. No similar relationship could be established between clinical parameters and A. actinomycetemcomitans. On a site level, the odds of detecting P. gingivalis increased by a factor of 2.47 (P= 0.0001) for every millimeter of residual probing depth; the odds of detecting P. intermedia/nigrescens increased by a factor of 1.84 (P= 0.0001). CONCLUSIONS: If, after standard mechanical periodontal therapy, a large number of sites continue to bleed, one may expect an increased number of sites positive for P. intermedia/ nigrescens. If many deep pockets persist, a greater number of P. gingivalis-positive sites can be expected.     

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.     

Changes in subgingival microflora and humoral immune response following periodontal therapy

OBJECTIVES: To investigate the effect of scaling and root planing (SRP) on the microflora and humoral immune response in adult periodontitis. MATERIALS & METHODS: Clinical measurements, subgingival plaque samples, gingival crevicular fluid and sera were taken from 4 sites in 28 adult periodontitis patients before and after SRP. Polymerase chain reaction was used to determine the presence of A. actinomycetemcomitans, P. gingivalis, B. forsythus, P. intermedia, and T. denticola. ELISA was used to investigate the systemic and local antibody titres to these organisms, and thiocyanate dissociation for the determination of serum antibody avidity. RESULTS: SRP produced a good clinical improvement. On a subject basis there was little significant change in the microflora. However, on a site basis, there were significant reductions in P. intermedia, B. forsythus and T. denticola. There was little change in systemic and local antibody titres following SRP, although there was a significant reduction in antibody avidity to P. gingivalis and P. intermedia CONCLUSION: Post-therapy clinical improvement was associated with a reduction in bacterial prevalence, but statistical significance was only reached at a site level and this microbial reduction was not significant for all organisms. No significant post-therapy effects on the humoral immune response were noted other than a reduced antibody avidity to P. gingivalis and P. intermedia. The lack of a clear pattern in the humoral immune response may reflect a failure of the host response to produce adequate levels of biologically functional antibodies, and complex interactions between the subgingival flora and the host response.     

牙周基础治疗对慢性牙周炎患者临床指标及5种牙周可疑致病微生物的影响

目的:观察牙周基础治疗对临床指标及5种牙周可疑致病微生物的影响。方法:选取20例慢性牙周炎患者(40个位点),在治疗前和基础治疗后3个月时检测观测位点的临床指标牙周探诊深度(PPD),临床附着丧失(CAL)和探诊出血(BOP),同时采集龈下微生物样本。采用PCR和反杂交的方法对所采集微生物样本中的牙龈卟啉单胞菌、福赛斯坦纳菌,中间普氏菌、伴放线放线杆菌和齿垢密螺旋体进行半定量检测。结果:通过牙周基础治疗后临床指标PPD及BOP的改善具有统计学意义(P<0.001),而CAL的改善不具有统计学意义。治疗后牙龈卟啉单胞菌、福赛斯坦纳菌和齿垢密螺旋体的检出量显著减少(P<0.05或P<0.001)。治疗前PPD>6mm的位点只有福赛斯坦纳菌在治疗后比治疗前有显著减少(P<0.05),而牙龈卟啉单胞菌和齿垢密螺旋体的变化不具有统计学意义。结论:基础治疗是治疗慢性牙周炎的有效方法,可改善临床指标,减少龈下牙龈卟啉单胞菌、福赛斯坦纳菌和齿垢密螺旋体的数量。但在PPD>6mm的位点基础治疗对于这五微生物的影响作用是有限的。     

Subgingival microbiota of chilean patients with chronic periodontitis

BACKGROUND: An association between race/ethnicity and the composition of the subgingival microbiota has been found in chronic periodontitis. A study was undertaken to determine the characteristics of the subgingival microbiota of chronic periodontitis in Chileans residing in Santiago. METHODS: Twenty-six subjects (mean age 45 +/- 7 years) with chronic periodontitis, mean probing depth (PD) 2.63 +/- 0.5 mm, mean attachment level (AL) 3.70 +/- 0.77 mm, and without a history of periodontal therapy were selected. Measurements of PD, AL, bleeding on probing, and plaque accumulation were recorded at six sites per tooth. Subgingival plaque samples were taken from the mesial aspect of every tooth and evaluated for the presence, levels, and proportions of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The microbial data of the Chileans were compared with data from 115 chronic periodontitis patients from Boston, Massachusetts. Since several clinical and demographic parameters differed between the two populations, significance of differences for each species was determined using analysis of covariance, adjusting for age, plaque level, mean PD, gender, and smoking status. RESULTS: Each of the individual test species was present in at least 25 of the 26 subjects, and 12 subjects (46.1%) harbored all 40 test species. With the exception of Prevotella intermedia, all test species colonized more than 75% of sites, and 25 species colonized > or = 90% of sites including the co-colonizing species of advanced periodontal lesions, termed the red complex, composed of the three species Porphyromonas gingivalis, Tannerella forsythensis (formerly Bacteroides forsythus), and Treponema denticola as well as Fusobacterium nucleatum subspecies, Campylobacter rectus, Peptostreptococcus micros, and Treponerma socranskii. Sixteen of the 40 species differed significantly between Chilean and U.S. subjects. Red, yellow, and other complexes were significantly higher in the Chileans, while the Actinomyces were higher in the U.S. subjects. CONCLUSIONS: The composition of the subgingival plaque differs among different subject populations. Thus, care should be taken when extrapolating the findings of one study to different ethnic groups.     

Multicenter evaluation of tetracycline fiber therapy. III. Microbiological response

In a multicenter study of the effects of tetracycline (TC) fiber therapy, subgingival plaque samples were tested for 6 probable periodontal pathogens by DNA probe analysis. Levels of Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas (Bacteroides) gingivalis, Prevotella intermedia (Bacteroides intermedius), and Wolinella recta were quantitatively determined in samples taken at baseline, and immediately after TC fiber removal, control fiber removal, and scaling and root planing. At untreated sites, samples were taken at baseline and 10 d later. Specificity of the DNA probe method was evaluated by testing the hybridization to 83 reference cultures. Interaction of the F. nucleatum probe with Fusobacterium periodonticum, and of the W. recta probe with Wolinella curva were the only cross-hybridizations noted. Species were detected at an average sensitivity of 2.9 x 10(4) organisms per sample. Approximately 70% of sites were initially infected with P. gingivalis and F. nucleatum; 50% with P. intermedia and E. corrodens; infections with W. recta and A. actinomycetemcomitans were less common (36% and 11% respectively). The average numbers of organisms found in the plaque samples were highest for F. nucleatum, P. gingivalis, and P. intermedia (ca. 10(6)). E. corrodens, W. recta, and A. actinomycetemcomitans occurred at 10-fold lower levels. Bacterial numbers and proportions of species in subgingival sites from the five centers did not differ appreciably. Both TC fiber therapy and scaling decreased the number of sites infected with all the monitored species. The bacterial composition at untreated sites and at sites where control fibers were placed was not significantly altered. The percentage reduction of the number of sites with detectable infection varied with each species: from 86% with W. recta to approximately 40% with P. gingivalis. Significant reduction of pocket depth and bleeding occurred at TC fiber-treated sites infected with each of the species. Significant attachment level gain occurred only at sites initially infected with P. gingivalis and treated with TC fibers.     

Comparison of the microbiota of supra- and subgingival plaque in health and periodontitis

BACKGROUND, AIMS: The purpose of the present investigation was to compare the microbial composition of supra and subgingival plaque in 22 periodontally healthy (mean age 32+/-16 years) and 23 adult periodontitis subjects (mean age 51+/-14 years). METHODS: A total of 2358 supra and separately subgingival plaque samples were collected from the mesial aspect of all teeth excluding 3rd molars in each subject. Samples were examined for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. Mean counts (x10(5), % DNA probe count and % sites colonized for each species were determined separately for supra and subgingival samples in each subject and then averaged across subjects in the 2 clinical groups. Significance of differences between healthy and periodontitis subjects was determined using the Mann-Whitney test and adjusted for multiple comparisons. RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) for healthy and periodontitis subjects in supragingival plaque were 72.1+/-11 and 132+/-17.5, respectively (p<0.01), and in subgingival plaque 22.1+/-6.6 and 100.3+/-18.4, (p<0.001). Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola could be detected in supragingival plaque samples of both healthy and periodontitis subjects. Actinomyces species were the dominant taxa in both supra- and subgingival plaque from healthy and periodontitis subjects. 4 Actinomyces species accounted for 63.2%, of supragingival and 47.2% of subgingival plaque in healthy subjects and 48.% and 37.8% in periodontitis subjects respectively. Increased proportions of P. gingivalis, B. forsythus, and species of Prevotella, Fusobacterium, Campylobacter and Treponema were detected subgingivally in the periodontitis subjects. P. gingivalis, B. forsythus and T. denticola were significantly more prevalent in both supra- and subgingival plaque samples from periodontitis subjects. CONCLUSIONS: The main differences between supra and subgingival plaque as well as between health and disease were in the proportions and to some extent levels of Actinomyces, "orange" and "red" complex species.     

Distribution of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in an Australian population

BACKGROUND, AIM: The present study describes (i) the natural distribution of the three putative periodontopathogens Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans in an Australian population and (ii) the relationship between these organisms, pocket depths and supragingival plaque scores. METHODS: Subgingival plaque was collected from the shallowest and deepest probing site in each sextant of the dentition. In total, 6030 subgingival plaque samples were collected from 504 subjects. An ELISA utilising pathogen-specific monoclonal antibodies was used to quantitate bacterial numbers. RESULTS:: A. actinomycetemcomitans was the most frequently detected organism (22.8% of subjects) followed by P. gingivalis and P. intermedia (14.7% and 9.5% of subjects respectively). The majority of infected subjects (83%) were colonised by a single species of organism. A. actinomycetemcomitans presence was over-represented in the youngest age group but under-represented in the older age groups. Conversely, P. gingivalis and P. intermedia presence was under-represented in the youngest age group but over-represented in the older age groups. Differing trends in the distribution of these bacteria were observed between subjects depending upon the site of the infection or whether a single or mixed infection was present; however, these differences did not reach significance. Bacterial presence was strongly associated with pocket depth for both A. actinomycetemcomitans and P. gingivalis. For A. actinomycetemcomitans, the odds of a site containing this bacterium decrease with deeper pockets. In contrast, for P. gingivalis the odds of a site being positive are almost six times greater for pockets >3 mm than for pockets < or =3 mm. These odds increase further to 15.3 for pockets deeper than 5 mm. The odds of a site being P. intermedia positive were marginally greater (1.16) for pockets deeper than 3 mm. CONCLUSIONS: This cross-sectional study in a volunteer Australian population, demonstrated recognised periodontal pathogens occur as part of the flora of the subgingival plaque. Prospective longitudinal studies are needed to examine the positive relationship between pocket depth and pathogen presence with periodontal disease initiation and/or progression.     

Comparison of subgingival bacterial sampling with oral lavage for detection and quantification of periodontal pathogens by real-time polymerase chain reaction

BACKGROUND: Saliva has been studied for the presence of subgingival pathogens in periodontitis patients. With the anaerobic culture technique, the discrepancy between salivary recovery and subgingival presence has been significant, which makes this approach not suitable for practical use in the microbial diagnosis of periodontitis patients. The real-time polymerase chain reaction (PCR) technique represents a very sensitive technique to detect and quantify bacterial pathogens. The aim of the study was to compare the presence and numbers of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia, and Micromonas micros in subgingival plaque and mouthwash samples by the anaerobic culture and real-time PCR techniques. METHODS: Pooled subgingival plaque samples and 10-ml mouthwash samples were collected from 21 adult patients with periodontitis and analyzed by quantitative anaerobic culture and real-time PCR for A. actinomycetemcomitans, P. gingivalis, T. forsythensis, P. intermedia, and M. micros. RESULTS: The detection frequency of A. actinomycetemcomitans, P. gingivalis, and T. forsythensis in subgingival plaque was identical by culture and real-time PCR and was higher for P. intermedia and M. micros by real-time PCR. The highest detection frequencies for the target bacteria were found in mouthwash samples by real-time PCR. The additional value of the real-time PCR to detect target bacteria was 38% for P. gingivalis, 73% for T. forsythensis, 77% for P. intermedia, and 71% for M. micros. The sensitivity to detect target species in mouthwash by real-time PCR was 100% for all test species except for P. intermedia (93.8%). CONCLUSIONS: Rapid detection and quantification of periodontal pathogens in mouthwash samples are possible by real-time PCR. The procedure is significantly less time-consuming than subgingival sampling with paper points. This approach to detect major periodontal pathogens in mouthwash samples may simplify microbial diagnosis in periodontitis patients and may be used to monitor periodontal treatment.     

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.     

Comparison of microbial cultivation and a commercial PCR based method for detection of periodontopathogenic species in subgingival plaque samples

OBJECTIVES: Microbiological laboratory procedures are involved in diagnosis and therapy control of progressive and refractory forms of periodontitis. In recent years techniques have been developed based on the detection of nucleic acids. The purpose of this study was to validate the commercially available micro-Dent(R) test which employs probes for A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus and T. denticola. METHODS: 122 plaque samples obtained from periodontal pockets with various depths from 33 early onset periodontitis (EOP) patients and 15 periodontally healthy subjects were analysed by cultivation and the microDent(R) kit. RESULTS: Both cultivation and the nucleic acid based assay showed a positive correlation of pocket depth with the frequency and quantity of periodontopathogenic species. T. denticola was found only in pockets > 4 mm in EOP patients. Comparison of the two methods revealed that the microDent(R) kit identified both P. gingivalis and B. forsythus more often than did the cultivation method. Conclusions: Nucleic acid techniques should replace cultivation methods as gold standard in microbiological diagnosis of progressive periodontitis. The micro-Dent(R) kit can be recommended for microbiological laboratories analysing subgingival plaque samples.     

An analysis of the subgingival microflora in randomly selected subjects

The aim of this study was to describe the presence of some microbial species in the subgingival microflora of a randomly selected subject sample. A further objective was to analyze the association between some microbiological characteristics and the periodontal conditions of the subjects. A total of 171 individuals aged 30–65 years were included. A pooled subgingival plaque sample was obtained from the mesial aspect of the 6 Ramfjord teeth by the use of the paper-point method. The samples were processed and analyzed according to standardized procedures. The periodontal condition of the subjects was examined clinically and included assessment of plaque, gingivitis, probing pocket depth and probing attachment level. The results showed that 81% of the subjects were positive for Campylobacter rectus , 80% for Fusobacterium nucleatum , 77% for Streptococcus sanguis , 72% for Streptococcus mitis , 66% for Eikenella corrodens , 58% for Prevotella intermedia , 32% for Capnocytophaga , 21% for Selenomonas , 25% for Actinobacillus actinomycetemcomitans , 14% for Porphyromonas gingivalis and 13% for Streptococcus mutans. Small differences in the periodontal conditions were observed between subjects harboring P. gingivalis, A. actinomytecomitans or >5% P. intermedia and the rest of the sample. Similarly small differences in the clinical parameters were revealed when the subjects were grouped according to proportions of disease-associated and health-associated species in their subgingival samples.     

Determination of eight selected periodontal pathogens in the subgingival plaque of maxillary first molars in Japanese school children aged 8-11 years

To prevent the onset or progression of periodontitis, we must understand when periodontal pathogens are first harbored and how they develop the biofilm that causes periodontal disease. The purpose of this study was to determine the relationship between clinical status and selected periodontal pathogens in subgingival plaque in school children. This study was conducted with 95 school children, 8-11 years old. The presence and distribution of eight selected periodontal pathogens sampled from the maxillary right first molar were determined by an indirect immunofluorescent technique and compared with clinical parameters. Of the 95 sites sampled, only one site had all eight pathogens and five sites did not have any of the eight pathogens. The mean number of positive pathogens per site was 3.5 +/- 1.8 and mean percentage of positive pathogens was 3.82 +/- 4.22%. The number and total percentage of positive pathogens were strongly correlated with the Plaque Index (PI). In addition, the number of positive pathogens was correlated to the presence of subgingival calculus. The most frequently found pathogens were Campylobacter rectus (84.2%) and Eikenella corrodens (83.2%), and the least, Actinobacillus actinomycetemcomitans serotype c (7.4%). Of the eight pathogens, the frequency and distribution of Porphyromonas gingivalis were significantly correlated with PI and the presence of calculus. In addition, seven sites with both P. gingivalis and Bacteroides forsythus showed a correlation with gingival inflammation. In conclusion, the presence of P. gingivalis or P. gingivalis and B. forsythus may be a risk marker to be sought in screening for the onset of periodontal disease.