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Journal ArticleDOI

Microbiological diagnosis of the severe chronic periodontitis

26 Mar 2010-Iss: 2009, pp 89-94

TL;DR: The presence of periodontopathogens as well as other bacterial species of possible importance should be considered in the patients with severe chronic periodontitis.

AbstractIn total, 14 adult patients with severe chronic periodontitis were evaluated for the presence of associated anaerobic and aerobic bacteria. Subgingival plaque specimens from three pocket depths per patient were obtained. Microaerophilic and facultative anaerobic bacteria, probably involved in the periodontitis, were isolated in six (42.9%) patients. These were Gram negative species involving Aggregatibacter (Haemophilus) aphrophilus (14.3%), Haemophilus parainfluenzae (7.1%), Kingella denitrificans (7.1%) and Moraxella osloensis (7.1%) as well as Gram-positive species, including Arcanobacterium (Actinomyces) pyogenes (7.1%) and Rhodococcus equi (7.1%). Anaerobic microbiology was completed for 12 patients. Of them, suspected periodontopathogens were isolated in seven (58.3%) patients and comprised Prevotella intermedia (in 41.7% of the patients) and Porphyromonas gingivalis (25%) as well as Porphyromonas endodontalis (8.3%). Tannerella forsythia was detected by PCR in half of the 12 cases. In conclusion, the presence of periodontopathogens as well as other bacterial species of possible importance should be considered in the patients with severe chronic periodontitis.

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Citations
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Journal ArticleDOI
TL;DR: This review examines literature data concerning the bacterial findings in chronic periodontitis depending on pocket depth, and presents the latest published information on the presence of proinflammatory factors in periodontal environment.
Abstract: This review examines literature data concerning the bacterial findings in chronic periodontitis depending on pocket depth, and presents the latest published information on the presence of proinflammatory factors in periodontal environment. It has been found that chronic periodontitis affects as much as 80% of the middle-aged population; by comparison, the prevalence of aggressive periodontitis reaches up to 1-1.5%. It is accepted that this social disease is multifactorial in etiology, but the evidence in the literature suggests that the levels of specific Gram-negative organisms in subgingival plaque biofilm play a major role in the initiation and progression of the disease. Of the many bacterial species inhabiting the periodontal environment, three types--Porphyromonas gingivalis (PG), Treponema denticola (TD), Tannerella forsythia (TF)--are strongly associated with the initiation and progression of periodontitis. Microbiological studies suggest that Porphyromonas gingivalis should be considered a major etiologic agent. Currently, Porphyromonas gingivalis is strongly associated with the pathogenesis of chronic periodontitis. On the other hand, the presence of Aggregatibacter actinomycetemeomitans in patients with chronic periodontitis may be related to the severity of the disease and thus modify the therapeutic plan. The increased amount of periodontal pathogens in the subgingival area can activate a cascade of defense mechanisms of the body associated with the production of factors causing inflammation and destruction, which suggests a correlation between the bacterial findings and the body response implemented by enhancing the local cytokine expression. Studies in the literature show that the presence of certain micro-organisms in the periodontal environment is associated to increased levels of proinflammatory cytokines in the gingival fluid and gingival tissue. These levels have been associated with destructive tissues response. There is little evidence in the literature on the correlation of the levels of periodontal pathogens of sites with different pocket depth with periodontal disease activity defined by the degree of the proinflammatory cytokine expression such as tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6).

29 citations


Journal ArticleDOI
TL;DR: This study investigated the detection rate of Porphyromonas gingivalis in chronic periodontitis patients versus healthy subjects using PCR assay, and its association with increased pocket depth and clinical attachment loss.
Abstract: Background and objective: Chronic periodontitis is the destruction of the tooth supporting structures as a result of a complex interaction between bacteria colonizing the gingival crevice and host’s immune responses. Porphyromonas gingivalis is one of the main periodontopathogens with multiple virulence factors. The aim of this study was to investigate the detection rate of Porphyromonas gingivalis in chronic periodontitis patients versus healthy subjects using PCR assay, and its association with increased pocket depth and clinical attachment loss. Methods: Seventy subjects (35 patients with chronic periodontitis and 35 healthy subjects) meeting the inclusion criteria of this study were selected. All the subjects were clinically assessed for probing pocket depth and clinical attachment loss then subgingival microbial samples were collected using sterile paper points and analyzed for the presence of Porphyromonas gingivalis using polymerase chain reaction assay. Results: A significant difference in Porphyromonas gingivalis detection rate between chronic periodontitis and healthy groups was recorded. Porphyromonas gingivalis was significantly associated with deep pockets. The detection rate increased with the increase in the severity of the disease, although, this correlation was not statistically significant. Conclusion: A positive association was observed between Porphyromonas gingivalis and increased pocket depth. The recovery rate was higher in severe cases.

3 citations


Journal ArticleDOI
01 Dec 2014
TL;DR: In this study high prevalence of anaerobic bacteria in the periodontal pockets of patients suffering from periodontitis compared with the gingival sulcus of healthy subjects with marked shifting from mainly Gram positive facultative anaerilic bacteria in shallow pockets to mainly Gram-negative strictAnaerobicacteria in the deep pockets.
Abstract: Aims: : The study evaluated the prevalence of anaerobic bacteria in the subgingival plaque in periodontitis in relation to pocket depth. Materials and Methods :The study was performed on 97 sub-gingival plague sample , pockets depth were measured , anaerobic bacteria were isolated and identified in relation to pockets depth. Results: In this study high prevalence of anaerobic bacteria in the periodontal pockets of patients suffering from periodontitis compared with the gingival sulcus of healthy subjects with marked shifting from mainly Gram positive facultative anaerobic bacteria in shallow pockets to mainly Gram-negative strict anaerobic bacteria in the deep pockets.Conclusion:. Anaerobic culture used in this study provided information about the susceptibility of the individuals to develop periodontal diseases. on bone.

3 citations


Cites background from "Microbiological diagnosis of the se..."

  • ...671) between pocket depth and the number of Gram-positive isolates.((15)) studied the microbiological diagnosis of the sever chronic periodontitis in which anaerobic microbiology was completed for 27 patients and found that Gram-negative anaerobic bacteria were isolated in 92....

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Journal ArticleDOI
07 Jul 2013
Abstract: 313 SUMMARY Periodontitis is an infectious disease concerning supporting tissues of the teeth. The primary etiological agent for disease development and progression is the subgingival biofilm, but recently it is known that host factors may modify the pathological process or may affect the severity and /or extent. The increasing levels of some specific pathogenic subgingival bacteria such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and others can result in periodontal destruction and possibly correlate with disease severity (3, 4, 5). Data from controlled studies show high prevalence of P. gingivalis, T. forsythia and Tr. denticola which represent the red complex (coexistence of these three species) in patients with moderate and severe chronic periodontitis (3, 5, 9). Parallel investigation of probing depth (PD) and clinical attachment level (CAL) with the microbiological testing may give a confirmation of relation between subgingival pathogenic bacteria and severity of periodontitis (1, 2, 7).

3 citations


Journal ArticleDOI
TL;DR: The study revealed that there are effects of age and sex on isolation rate and the results indicated that percentage of P.gingivalis was detected in 20-30 years old and males were more infected than females.
Abstract: The study aimed to detect P. gingivalis from 49 patients with periodontitis at different ages and both sexes, after determination of pocket depth, types of infection whether chronic or progressive by dentists. Routine culture method was done using selective media and anaerobic condition and compared with species specific polymerase chain reaction (PCR) technique. DNA was extracted from samples and its concentration and purity were determined. The results showed domination of chronic infections and the pocket depths ranged between 3-9mm, as well as the results revealed that isolation percent of P.gingivalis by PCR was more higher than culture method, it was 65.3% and 28.5% respectively. The results also showed that phenol-chloroform was the efficient method for DNA extraction comparing with other methods. The study revealed that there are effects of age and sex on isolation rate and the results indicated that percentage of P.gingivalis was detected in 20-30 years old and males were more infected than females.

2 citations


References
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Journal ArticleDOI
Abstract: Recent investigations of the human subgingival oral flora based on ribosomal 16S cloning and sequencing have shown many of the bacterial species present to be novel species or phylotypes. The purpose of the present investigation was to identify potential periodontal pathogens among these newly identified species and phylotypes. Species-specific ribosomal 16S primers for PCR amplification were developed for detection of new species. Associations with chronic periodontitis were observed for several new species or phylotypes, including uncultivated clones D084 and BH017 from the Deferribacteres phylum, AU126 from the Bacteroidetes phylum, Megasphaera clone BB166, clone X112 from the OP11 phylum, and clone I025 from the TM7 phylum, and the named species Eubacterium saphenum, Porphyromonas endodontalis, Prevotella denticola, and Cryptobacterium curtum. Species or phylotypes more prevalent in periodontal health included two uncultivated phylotypes, clone W090 from the Deferribacteres phylum and clone BU063 from the Bacteroidetes, and named species Atopobium rimae and Atopobium parvulum.

453 citations


Journal ArticleDOI
TL;DR: Chronic periodontitis is the result of a global perturbation of the oral bacterial ecology rather than a disease-site specific microbial shift, and more differences were found in the bacterial profile between subjects withperiodontitis and healthy subjects than between deep and shallow sites within the same subject.
Abstract: Most studies of the bacterial etiology of periodontitis have used either culture-based or targeted DNA approaches, and so it is likely that pathogens remain undiscovered. The purpose of this study was to use culture-independent, quantitative analysis of biofilms associated with chronic periodontitis and periodontal health to identify pathogens and beneficial species. Samples from subjects with periodontitis and controls were analyzed using ribosomal 16S cloning and sequencing. Several genera, many of them uncultivated, were associated with periodontitis, the most numerous of which were gram positive, including Peptostreptococcus and Filifactor. The genera Megasphaera and Desulfobulbus were elevated in periodontitis, and the levels of several species or phylotypes of Campylobacter, Selenomonas, Deferribacteres, Dialister, Catonella, Tannerella, Streptococcus, Atopobium, Eubacterium, and Treponema were elevated in disease. Streptococcus and Veillonella spp. were found in high numbers in all samples and accounted for a significantly greater fraction of the microbial community in healthy subjects than in those with periodontitis. The microbial profile of periodontal health also included the less-abundant genera Campylobacter, Abiotrophia, Gemella, Capnocytophaga, and Neisseria. These newly identified candidates outnumbered Porphyromonas gingivalis and other species previously implicated as periodontopathogens, and it is not clear if newly identified and more numerous species may play a more important role in pathogenesis. Finally, more differences were found in the bacterial profile between subjects with periodontitis and healthy subjects than between deep and shallow sites within the same subject. This suggests that chronic periodontitis is the result of a global perturbation of the oral bacterial ecology rather than a disease-site specific microbial shift.

414 citations


Journal ArticleDOI
TL;DR: The present findings using a large-scale analysis allowed the inclusion of some newly named species and as-yet-uncultivated phylotypes in the set of candidate pathogens associated with this disease.
Abstract: Samples from infected root canals of 43 teeth with chronic apical periodontitis were analyzed for the presence and relative levels of 83 oral bacterial species and/or phylotypes using a reverse-capture checkerboard hybridization assay. Associations between the most frequently detected taxa were also recorded. The most prevalent taxa were Olsenella uli (74%), Eikenella corrodens (63%), Porphyromonas endodontalis (56%), Peptostreptococcus anaerobius (54%), and Bacteroidetes oral clone X083 (51%). When prevalence was considered only for bacteria present at levels >10(5), Bacteroidetes clone X083 was the most frequently isolated bacterium (37%), followed by Parvimonas micra (28%), E. corrodens (23%), and Tannerella forsythia (19%). The number of target taxa per canal was directly proportional to the size of the apical periodontitis lesion, with lesions >10 mm in diameter harboring a mean number of approximately 20 taxa. Several positive associations for the most prevalent taxa were disclosed for the first time and may have important ecological and pathogenic implications. In addition to strengthening the association of several cultivable named species with chronic apical periodontitis, the present findings using a large-scale analysis allowed the inclusion of some newly named species and as-yet-uncultivated phylotypes in the set of candidate pathogens associated with this disease.

133 citations


Journal ArticleDOI
TL;DR: Rapid detection and quantification of periodontal pathogens in mouthwash samples are possible by real-time PCR, which is significantly less time-consuming than subgingival sampling with paper points.
Abstract: 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.

117 citations


Additional excerpts

  • ...DISCUSSION Boutaga et al. (2007) have reported the real-time polymerase chain reaction (PCR) to be a very sensitive technique to detect bacterial periodontopathogens....

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Journal ArticleDOI
TL;DR: It is demonstrated that a wide range of oral species may be detected on or in crevicular epithelial cells from sites with periodontitis and from periodontally healthy sulci.
Abstract: Bacterial invasion of host epithelial cells plays an important role in the pathogenesis of periodontal diseases; however, the interactions between subgingival species and the gingival crevice cells are not fully understood. This study determined the prevalence of a group of oral bacterial species on or in epithelial cells derived from periodontal pockets and the gingival crevice of subjects with periodontitis. Samples of epithelial cells were obtained from 120 sites with periodontal pockets > or =4 mm and 92 periodontally healthy sites from 49 patients (mean age 46.3+/-1.4 years; 43% males) with chronic periodontitis. Bacteria in or on epithelial cells were separated from unattached bacteria by Percoll density-gradient centrifugation. The presence and levels of 33 oral species were determined in epithelial cell samples by whole genomic DNA probes and the checkerboard method. The most frequently detected species were Porphyromonas gingivalis (42%), Treponema denticola (38%), Prevotella intermedia (37%), Streptococcus intermedius (36%), Campylobacter rectus (35%), Streptococcus sanguinis (35%) and Streptococcus oralis (34%). Species of Actinomyces were found in low prevalence and levels. The data indicated that there were more micro-organisms on or in epithelial cells obtained from periodontal pockets than from healthy sulci; however, no significant differences regarding the percentage and level of any specific species were found between these sites. Veillonella parvula, S. oralis, Streptococcus gordonii and Streptococcus mitis tended to be more prevalent in sites without disease. These findings demonstrated that a wide range of oral species may be detected on or in crevicular epithelial cells from sites with periodontitis and from periodontally healthy sulci.

95 citations