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.     

Acquisition and loss of Porphyromonas gingivalis,Actinobacillus actinomycetemcomitans and Prevotella intermedia over a 5-year period: effect of a triclosan/copolymer dentifrice

OBJECTIVES: The present study describes the natural history of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia over a 5-year period and the effect of a triclosan/copolymer dentifrice on these organisms in a normal adult population. MATERIAL AND METHODS: Subgingival plaque samples were collected from 504 adult volunteers. Probing pocket depths (PPD) and relative attachment levels were measured using an automated probe. Participants were matched for disease status (CPI), plaque index, age and gender, and allocated to receive either a triclosan/copolymer or placebo dentifrice. Re-examination and subgingival plaque sampling was repeated after 1, 2, 3, 4 and 5 years. P. gingivalis, A. actinomycetemcomitans and P. intermedia were detected and quantitated using an enzyme linked immunosorbent assay. Logistic regression and generalised linear modelling were used to analyse the data. RESULTS: This 5-year longitudinal study showed considerable volatility in acquisition and loss (below the level of detection) of all three organisms in this population. Relatively few subjects had these organisms on multiple occasions. While P. gingivalis was related to loss of attachment and to PPD >/=3.5 mm, there was no relationship between A. actinomycetemcomitans or P. intermedia and disease progression over the 5 years of the study. Smokers with P. gingivalis had more PPD >/=3.5 mm than smokers without this organism. There was no significant effect of the triclosan dentifrice on P. gingivalis or A. actinomycetemcomitans. Subjects using triclosan were more likely to have P. intermedia than those not using the dentifrice; however this did not translate into these subjects having higher levels of P. intermedia and its presence was uniform showing no signs of increasing over the course of the study. CONCLUSION: The present 5-year longitudinal study has shown the transient nature of colonisation with P. gingivalis, A. actinomycetemcomitans and P. intermedia in a normal adult population. The use of a triclosan-containing dentifrice did not lead to an overgrowth of these organisms. The clinical effect of the dentifrice would appear to be independent of its antimicrobial properties.     

Microbial associations in periodontitis sites before and after treatment

Duplicate samples from 110 periodontal sites of 6 mm or more pocket depth in 16 patients were analyzed for the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Capnocytophaga spp., Campylobacter rectus, Eikenella corrodens and Fusobacterium nucleatum. The sites were sampled before and after nonsurgical periodontal treatment. No statistically significant associations were found before treatment between any of the analyzed species. After treatment, statistically significant associations were found between E. corrodens and all the other species, F. nucleatum and P. intermedia; Capnocytophaga spp. and C. rectus; P. intermedia vs Capnocytophaga spp. and P. gingivalis ; and C. rectus vs Capnocytophaga spp. and A. actinomycetemcomitans. Some of these associations could be explained either by patient-related factors or site-related characteristics such as the pocket depth. The proportion of P. gingivalis seemed to be unrelated to the proportion of P. intermedia in the samples. If one of the analyzed microbes was found in one of the sampled pockets in a patient, the probability of finding that microbe in all the sampled sites in the same patient before treatment was more than 50%. This probability was reduced after treatment for many species, especially P. gingivalis , which showed a probability of zero. The probability of detecting a bacterial species on at least one additional site if it was present on one in the same individual was nearly 100%, both before and after treatment, for all species studied. This study has shown several potential microbial associations in the subgingival plaque flora of deep periodontal pockets. Local factors of the periodontal pocket and factors related to the individual may mask the biological significance of these microbial interactions.     

Actinobacillus actinomycetemcomitans, Capnocytophaga and Porphyromonas gingivalis in subgingival plaque of adolescents with Down''s syndrome

Levels of Actinobacillus actinomycetemcomitans, Capnocytophaga and Porphyromonas gingivalis were determined in subgingival plaque samples from 37 adolescents with Down's syndrome and 37 healthy controls matched with respect to age and sex. Gingival inflammation, supra- and subgingival calculus, periodontal pockets ( > 4 mm) and alveolar bone loss were registered. Alveolar bone loss was more frequent in Down's syndrome subjects (32%) than in the controls (3%). A. actinomycetemcomitans was detected in the subgingival plaque in 35% of the Down's syndrome adolescents and in 5% of the controls. On site level, A. actinomycetemcomitans and Capnocytophaga were more frequent in the subgingival plaque samples of Down's syndrome children than in those of controls. Comparing Down's syndrome subjects positive or negative for A. actinomycetemcomitans and Capnocytophaga, no significant differences were found in terms of gingival inflammation, periodontal pockets ( > 4 mm) or number of sites with alveolar bone loss. The results indicate an altered microbial composition of the subgingival plaque of Down's syndrome subjects compared with healthy controls, with higher frequency of A. actinomycetemcomitans.     

DNA probe detection of periodontopathogens in advanced periodontitis

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 seein to be important in the pathogenesis of the disease.     

Untreated periodontal disease in Indonesian adolescents. Longitudinal clinical data and prospective clinical and microbiological risk assessment

BACKGROUND, AIMS: In order to investigate the r le of various putative clinical and microbiological risk markers, a longitudinal study was initiated in a young population deprived of regular dental care. In 1987 all inhabitants in the age range 15-25 years living in a village with approximately 2,000 inhabitants at a tea estate on Western Java, Indonesia, were examined clinically and microbiologically. In total, 167 subjects of the original group of 255 adolescents were re-examined in 1994. The material presented in this paper describes the clinical periodontal condition at baseline (1987) and at follow-up (1994), 7 years later. Furthermore, the relationship between progression of the disease and baseline clinical and microbiological data was assessed. METHODS: Plaque index (PI), bleeding on probing (BOP), pocket depth (PD), and attachment loss (AL) were scored at the approximal surfaces of the vestibular aspects of all teeth. The number of approximal surfaces of the Ramfjord teeth showing subgingival calculus was recorded. At baseline, the dorsum of the tongue, the buccal gingiva in the upper jaw, the saliva and the deepest bleeding pocket without clinical loss of attachment were sampled for microbiological examination with phase contrast microscopy and indirect immunofluorescence. RESULTS: Mean values at baseline and at follow-up were PI: 1.01 and 1.15, BOP: 0.80 and 1.16, PD 3.26 mm and 3.32 mm, AL: 0.33 mm and 0.73 mm, respectively. All parameters except PD showed a statistically significant increase over the 7-year period. The prevalence of the studied bacteria irrespective of the sample site was: A. actinomycetemcomitans 53%, P. gingivalis 88%, P. intermedia 100%, spirochetes 89% and motile micro-organisms 100%. At the full mouth level, logistic regression showed significant odds ratios for progressive disease with age (1.15), subgingival calculus (1.20) and subgingival presence of A. actinomycetemcomitans (4.61). Presence of any of the selected micro-organisms on the mucous membranes was not related with progressive disease. In order to study local factors to explain local disease activity, each subject was characterized using the sampled pocket, which was the deepest bleeding pocket without LA at baseline, as a single response site per patient. In this constrained design, the main statistical factors associated with progressive disease were presence of motile micro-organisms and the plaque score. CONCLUSIONS: This study identified 3 main risk markers for disease progression at the full mouth level: age, amount of subgingival calculus and subgingival presence of A. actinomycetemcomitans.     

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.     

Persistent bacterial colonization of Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans in periodontitis and its association with alveolar bone loss after 6 months of therapy

BACKGROUND, AIMS: The purpose of this study was to determine whether the presence of bacterial antigens for Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), and Actinobacillus actinomycetemcomitans (Aa) in subgingival plaque of periodontitis patients after periodontal treatment was associated with progressive alveolar bone loss. METHOD: 39 (39) subjects in good general health previously diagnosed with adult periodontitis within the last 2 years, and still presenting with probing depth >5 mm in 2 to 6 teeth, were studied. All subjects were treated with scaling and root planing. Half of the subjects were randomly assigned to receive adjunctive systemic doxycycline (200 mg the 1st day, then 100 mg per day for 21 days). Subgingival plaque samples were taken at baseline, 1, 3 and 6 months after therapy. A modified ELISA test (Evalusite, Periodontal Test Kit, Eastman Kodak Co., Rochester, NY) was used to test for plaque antigens associated with P. gingivalis, P. intermedia and A. actinomycetemcomitans. Progressive alveolar bone loss was determined using digital subtraction radiography with standardized radiographs taken at baseline and 6 months after treatment. RESULTS: The presence of P. gingivalis in plaque after treatment was significantly associated with progressive bone loss (positive predictive value 84%, negative predictive value 85%, odds ratio 31.9, p<0.0001). In contrast, the presence of P. intermedia in plaque after treatment was not indicative of progressive loss (positive predictive value 39%, negative predictive value 82%). Too few sites had evidence of A. actinomycetemcomitans to be amenable to statistical analysis. No significant difference in bone loss was attributable to the systemic antibiotic therapy. CONCLUSION: These data indicated that, in this population, the presence of P. gingivalis in plaque after treatment might be indicative of progressive alveolar bone loss.     

Java project on periodontal diseases. The natural development of periodontitis: risk factors, risk predictors and risk determinants

OBJECTIVE: To identify risk factors, risk predictors and risk determinants for onset and progression of periodontitis. MATERIAL AND METHODS: For this longitudinal, prospective study all subjects in the age range 15-25 years living in a village of approximately 2000 inhabitants at a tea estate on Western Java, Indonesia, were selected. Baseline examination was carried out in 1987 and follow-up examinations in 1994 and 2002. In 2002, 128 subjects could be retrieved from the original group of 255. Baseline examination included evaluation of plaque, bleeding on probing, calculus, pocket depth, attachment loss and presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, spirochetes and motile microorganisms. RESULTS: The mean attachment loss increased from 0.33 mm in 1987 to 0.72 mm in 1994 and 1.97 mm in 2002. Analysis identified the amount of subgingival calculus and subgingival presence of A. actinomycetemcomitans as risk factors, and age as a risk determinant, for the onset of disease. Regarding disease progression, the number of sites with a probing depth > or = 5 mm and the number of sites with recession were identified as risk predictors and male gender as a risk determinant. CONCLUSION: Screening of these parameters early in life could be helpful in the prevention of onset and progression of periodontal diseases.     

Untreated periodontal disease in Indonesian adolescents

BACKGROUND/AIMS: In an Indonesian population deprived of regular dental care, the experienced progression of disease between baseline (1987) and follow-up (1994) was investigated in relation to the composition of the subgingival microbiota at follow-up. At baseline the age ranged from 15 to 25 years. Clinical and microbiological evaluation was completed in 158 of the 167 subjects available at follow-up. METHODS: Plaque index (PI), pocket depth (PD), bleeding on probing (BOP), and attachment loss (AL) were scored at the approximal surfaces of all teeth and subgingival calculus on the approximal surfaces of the Ramfjord teeth only (number of sites with subgingival calculus: NSC). A pooled sample of the deepest pocket in each quadrant was evaluated using microbiological culture techniques. RESULTS: At baseline the mean values of the clinical parameters were AL=0.35 mm, PI=1.01, BOP=0.80 PD=3.25 mm and NSC=6.04 and at follow-up AL=0.75 mm, PI=1.16, BOP=1.19, PD=3.34 mm and NSC=5.85. All parameters except PD and NSC showed a statistically significant increase. At follow-up the prevalence of Actinobacillus actinomycetemcomitans was 40%, of Porphyromonas gingivalis 67%, of Prevotella intermedia 66%, of Fusobacterium nucleatum 79%, of Bacteroides forsythus 16%, of Campylobacter rectus 4%, and of P. micros 6%. No differences in clinical parameters were found between groups with or without these micro-organisms. In 129 subjects AL of > or =2 mm at > or =1 site was found. Logistic regression showed three significant odds-ratio's for experienced progressive periodontitis: Plaque index (12.2), gender (3.4) and Actinobacillus actinomycetemcomitans (2.9). CONCLUSIONS: The results of this retrospective study suggest that plaque is the most important parameter related to experienced disease progression, and that the presence of A. actinomycetemcomitans may be associated with increased chance of disease progression.     

Evaluation of oral bacteria as risk indicators for periodontitis in older adults.

The prevalence of people and sites with attachment loss, pocket depth, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Porphyromonas gingivalis are described for a random sample of 366 black and 297 white community-dwelling adults, aged 65 or over, residing in five counties in North Carolina. In addition, relationships between sites harboring these microorganisms and loss of attachment (LA) and pocket depth (PD) are presented in a manner that considers the lack of independence of sites within each person. Pocket depths and recession were measured on all teeth by trained examiners during household visits. Immunofluorescent assays for A. actinomycetecomitans, P. intermedia, and P. gingivalis were conducted on subgingival plaque samples obtained from the mesiobuccal aspect of the four first molar teeth using paper points. The prevalences of A. actinomycetemcomitans, P. intermedia, and P. gingivalis were greater in blacks than in whites. The most striking difference was seen for P. gingivalis, which was found in 38.8% of blacks and 9.4% of whites. Similar relationships were found when the percent of sites with these organisms were assessed. Blacks with P. gingivalis or P. intermedia had a higher prevalence of sites with LA greater than or equal to 7 mm as compared to blacks not infected with P. gingivalis or P. intermedia. The same was true for whites. Similar relationships between P. gingivalis or P. intermedia and PD greater than or equal to 6 mm were found for both blacks and whites.(ABSTRACT TRUNCATED AT 250 WORDS)     

5种牙周龈下可疑致病微生物与慢性牙周炎局部不同牙周状态间的关系

目的 观察5种龈下微生物检出水平与慢性牙周炎局部牙周状态的关系。方法 选择20例慢性牙周炎患者的80个位点及10例牙周健康者的20个位点为观察位点,采集龈下微生物样本,记录牙周探诊深度（PD）,根据所测位点的PD进行分组。PD≤4 mm为A组,4 mm＜PD≤6 mm为B组,PD>6 mm为C组,健康对照组为H组。通过聚合酶链反应（PCR）和DNA探针反杂交技术半定量检测各组伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌的检出率和检出水平。结果 B、C组牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌的检出率和检出水平均高于H组,A组牙龈卟啉单胞菌的检出率和检出水平也高于H组,C组福赛斯坦纳菌和齿垢密螺旋体检出水平高于B组,以上差异均有统计学意义（P<0.05）;伴放线菌嗜血菌在各组间的检出率及检出水平都无明显差异。结论 随着牙周袋的加深,牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌体的阳性检出率和检出水平都有随之增加的趋势;牙龈卟啉单胞菌与慢性牙周炎的早期炎症关系较为密切,而福赛斯坦纳菌和齿垢密螺旋体与中重度慢性牙周炎炎症位点的严重程度有关。     

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.     

Real-time polymerase chain reaction for detection and quantification of bacteria in periodontal patients

BACKGROUND: Accurate laboratory tests for the detection and quantification of periodontopathogens in subgingival plaque samples of periodontal disease patients are becoming essential to study the pathogenesis of this polymicrobial condition. We used a real-time polymerase chain reaction (PCR) assay for the quantification of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Dialister pneumosintes, Campylobacter rectus, and Micromonas micros as well as total eubacteria in subgingival plaque samples from individuals with periodontitis. METHODS: Eighty-three subgingival samples from periodontally diseased patients and 43 samples from periodontally healthy subjects were tested and the results of bacterial quantification were correlated to clinical parameters. Quantification was performed with specific 16S rRNA target sequences with double fluorescence labeled probes and serial dilutions of plasmid standards by real-time PCR. RESULTS: Results showed that patients as well as healthy subjects were positive for the presence of target periodontopathogens; however, median values were higher in samples from periodontitis subjects. In addition, a positive association was observed between colonization at high levels by P. gingivalis and M. micros and the presence of deep periodontal pockets. CONCLUSION: Real-time PCR provides a reliable high-throughput method for quantification of periodontopathogens and may be useful for understanding the complex etiology observed in periodontal diseases.     

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.     

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.     

Microbiologic analysis of periodontal pockets and carotid atheromatous plaques in advanced chronic periodontitis patients

BACKGROUND: In recent years, many researchers have focused their attention on the ability of periodontal pathogens to colonize atheromatous plaques. Nevertheless, a clear correlation between the detection rates of periodontopathic bacterial DNA in atheromas and in subgingival plaque samples has not been established. The aim of our study was to assess the presence of five periodontal pathogens (Actinobacillus actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia [formerly Tannerella forsythensis]) in periodontal pockets and in carotid atheromas recovered from the same patients. METHODS: Thirty-three patients with advanced chronic periodontitis scheduled for endarterectomy were enrolled in the study. DNA was extracted from subgingival plaque samples and carotid atheromas. Universal bacteria primers for general detection of bacteria and species-specific primers for detection of periodontal pathogens were used to amplify part of the 16S rRNA gene by polymerase chain reaction. RESULTS: All subgingival plaque samples were positive for at least one target microorganism. The prevalence of T. forsythia, P. gingivalis, T. denticola, P. intermedia, and A. actinomycetemcomitans were 69.7%, 63.6%, 54.5%, 45.4%, and 33.3%, respectively. Bacterial DNA was detected in 31 out of 33 endarterectomy specimens. However, none of the samples tested positive for DNA from periodontal pathogens. CONCLUSION: The presence of periodontal bacteria in atheromatous plaques was not confirmed by this investigation; thus, no correlation could be drawn between periodontitis bacteria and microorganisms involved in the atherosclerotic lesions.     

慢性牙周炎龈下菌斑中五种牙周可疑致病微生物的分布

目的研究五种牙周可疑致病微生物在慢性牙周炎患者龈下菌斑的分布。方法选择27例慢性牙周炎患者,每位患者选取牙周袋最深的两个位点作为观察位点,采集龈下微生物样本,采用多重聚合酶链反应和反杂交的方法对伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体五种微生物进行半定量检测。结果在所检测的54个位点中,牙龈卟啉单胞菌、中间普雷沃菌、福赛斯坦纳菌和齿垢密螺旋体均有较高的检出率,分别为98.15%、92.59%、100%和98.15%;伴放线菌嗜血菌检出率较低,为20.37%。牙龈卟啉单胞菌和福赛斯坦纳菌的检出量明显高于其他三种微生物,其差异有统计学意义（P<0.05）。结论慢性牙周炎患者多存在牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体的同时感染,且牙龈卟啉单胞菌和福赛斯坦纳菌的感染量较高。     

The limited value of three pathogen species in predicting healing of periodontal pockets

Baseline level of Actinobacillus actinomycetemcomitans has been suggested as being predictive of periodontal treatment outcome. We analyzed the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in 55 deep periodontal pockets of 29 patients (18 men, 11 women, 37-75 years) before and after periodontal treatment. At baseline and after treatment, 62% and 33%, respectively, of the subjects presented with 1, 2, or a combination of all 3 pathogens. The mean pocket depth of 6.6 mm (0.4 mm) before treatment decreased to 2.2 mm (0.4 mm) in response to treatment (P<0.001). The treatment plan of non-surgical or surgical treatment was based on pocket depths and tooth morphology only. No antimicrobial medications were used during the treatment. Eighty-two percent of the deep pockets healed satisfactorily to < or = 4 mm. The presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, or Prevotella intermedia at baseline was not associated with the outcome of the periodontal therapy. In conclusion, we found that the presence of the 3 periodontopathogen species had little or no value in predicting healing of periodontal pockets.     

Microbiological profile of early onset/aggressive periodontitis patients

OBJECTIVES: The objectives of this study were to characterize the bacterial profile and to seek possible bacterial associations in the subgingival microbiota of early onset periodontitis/aggressive periodontitis patients by using two different techniques, culture and immunofluorescence. MATERIAL AND METHODS: The study group consisted of 66 systemically healthy individuals with evidence of early onset periodontitis - 41 females and 25 males aged 23-35 years (mean 31.1 +/- 3.1 years). Bacterial samples were collected from the deepest site in each quadrant, resulting in a total of 264 sites with a mean probing pocket depth of 6.6 +/- 1.5 mm. Samples were cultured anaerobically and in 10% CO(2) using selective and nonselective media, and isolates were characterized to species level. Indirect immunofluorescence using monoclonal antibodies was applied to detect Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia (Bacteroides forsythus, Tannerella forsythensis), Prevotella intermedia/Prevotella nigrescens, Campylobacter rectus, Peptostreptococcus micros and Actinomyces israelii. RESULTS: 93.6% of sampled sites showed bleeding on probing and 23.5% were positive for suppuration. P. intermedia/P. nigrescens, P. gingivalis, and C. rectus were detected in 77.3-85.9% of samples using culture methods and in 85.6-91.3% using immunofluorescence. P. micros and A. actinomycetemcomitans were found, respectively, in 63.3% and 25.0% of all sites using culturing and in 58.7% and 27.7% sites using immunofluorescence. Significantly strong positive associations were observed between T. forsythia and C. rectus (odds ratio 109.46), and T. forsythia and P. gingivalis (odd ratio 90.26), whereas a negative association was seen between P. intermedia/P. nigrescens and A. actinomycetemcomitans (odds ratio 0.42). Coinfection by P. gingivalis, T. forsythia, P. intermedia/P. nigrescens and C. rectus was observed in 62.1% of the test sites, and in 89.4% of the studied subjects. The sensitivity of immunofluorescence for T. forsythia, C. rectus, P. intermedia/P. nigrescens and P. gingivalis was found to be very high (0.99-0.94) using culture as the reference detection method. The agreement between culture and immunofluorescence in detecting the presence or absence of the investigated species was 85.2-88.1% for P. gingivalis, P. intermedia/P. nigrescens, C. rectus, and T. forsythia, 75.9% for A. actinomycetemcomitans and 70.4% for P. micros. CONCLUSIONS: The microbial profile of the early onset/aggressive periodontitis population was complex. The agreement between the two detection methods was very high.