Polymerase Chain Reaction of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in Primary Endodontic Infections (original) (raw)
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Oral Microbiology and Immunology, 2005
The aim of this study was to investigate the presence of four black-pigmented bacteria, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and Prevotella nigrescens, in endodontic infections by culture and polymerase chain reaction (PCR) analyses. Microbial samples were obtained from 50 teeth with untreated necrotic pulps (primary infection) and from 50 teeth with failing endodontic treatment (secondary infection). Microbiological strict anaerobic techniques were used for serial dilution, plating, incubation, and identification. For PCR detection, the samples were analyzed using species-specific primers of 16S rDNA and the downstream intergenic spacer region. Culture and PCR detected the test species in 13/100 and 50/100 of the study teeth, respectively. The organisms were cultured from 11/50 (22%) of primarily infected root canal samples and from 2/50 (4%) of secondary root canal samples. PCR detection identified the target species in 32/50 (64%) and 18/50 (36%) of primary and secondary infections, respectively. P. gingivalis was rarely isolated by culture methods (1%), but was the most frequently identified test species by PCR (38%). Similarly, P. endodontalis was not recovered by culture from any tooth studied, but was detected by PCR in 25% of the sampled teeth. PCR-based identification also showed higher detection rates of P. intermedia (33%) and P. nigrescens (22%) than culture (13%). In conclusion, P. gingivalis, P. endodontalis, P. intermedia, and P. nigrescens were identified more frequently in teeth with necrotic pulp than in teeth with failing endodontic treatment. Also, a higher frequency of black-pigmented species was detected by PCR than by culture.
Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 2007
Black-pigmented anaerobic rods such as Prevotella spp. and Porphyromonas spp. are involved in the etiology and perpetuation of endodontic infections. The aim of this study was to evaluate the prevalence of these species in chronic endodontic infections by using culture and polymerase chain reaction (PCR) techniques. Samples of 100 patients with root canals displaying chronic endodontic infections were obtained by sterilized paper points. Bacterial identification was performed by using culture and PCR techniques. By culture, in 33% of the samples, P. intermedia-P. nigrescens (75.8%), P. gingivalis (27.3%), and P. endodontalis (9.1%) were identified, and by PCR 60% of the samples harbored P. nigrescens (43.3%), P. gingivalis (43.3%), P. intermedia (31.7%), and P. endodontalis (23.3%). The presence of these black-pigmented anaerobic rods alone or in association in chronic endodontic infections seems to be frequent. PCR is a very sensitive technique for detecting DNA from bacterial cells. Culturing is only able to reveal living bacteria and is less sensitive for the identification of low numbers of bacterial cells.
African Journal of Microbiology Research, 2014
The aim of this investigation was to examine the presence of 8 bacterial anaerobic species in endodontic samples from patients with primary and secondary infection. The association of clinical signs and symptoms with constituent species were also evaluated. Microbial samples were obtained from 72 teeth with primary endodontic infection and 35 teeth with secondary endodontic infection. DNA was extracted from samples and analyzed with a polymerase chain reaction (PCR)-based identification assay. Medical and dental histories were obtained from each patient. The prevalence of the targeted bacterial species was recorded for each case and descriptive statistical analyses were performed using the Pearson Chi-squared test. Nucleid acid amplification method (NAAM) analysis showed that all specimens were positive at least for 1 or more samples in primary and secondary infection teeth. The most frequently detected bacteria in all speciemens were Porphyromonas gingivalis, followed by Porphyromonas micros, Porphyromonas endodontalis, Fusobacterium nucleatum, Porphyromonas intermedia and Tannerella forsythia, respectively. The percentages of all selected bacteria found in primary infection group were higher than secondary infection group except for Porphyromonas intermedia. However, statistically significant difference was found only for T. forsythia and F. nucleatum which were higher percentage in primary infection than in secondary infection group. There was a significant association between tenderness to percussion and P. gingivalis (p < 0.05), pain with Porphyromonas melaninogenica (p < 0.05) and swelling with both P. gingivalis (p < 0.05) and P. melaninogenica (p < 0.05). Findings indicated that the prevalence of some species found in the primary infection group were higher than in the secondary infection group. In this study there was a significant association between tenderness to percussion and P. gingivalis, pain with P. melaninogenica and swelling with both P. gingivalis and P. melaninogenica.
Journal of Endodontics, 2006
The aim of this study was to investigate the presence of strict anaerobes such as Filifactor alocis, Tannerella forsythia, and Treponema denticola in primary and secondary root-infected canals with periapical lesions by molecular analysis and the association of these species with specific endodontic signs and symptoms. Microbial samples were taken from 100 root canals, 50 with necrotic pulp tissues (NPT, primary infection), and 50 with failed endodontic treatment (FET, secondary infection). DNA was extracted from the samples, which were analyzed for the presence of three endodontic pathogens using species-specific primers and PCR. F. alocis were isolated from 23 canals with NPT and 12 canals with FET; T. forsythia from 12 canals with NPT and three canals with FET; T. denticola from 19 canals with NPT and 12 canals with TEP. Suggested associations were found between primary infection and the presence of F. alocis and T. forsythia (both p < 0.05). In particular, associations were found between: pain and F. alocis; swelling and F. alocis; tenderness to percussion and T. forsythia; mobility and T. forsythia and T. denticola; wet canals and F. alocis, T. forsythia, and T. denticola; purulent exsudate and F. alocis, T. forsythia and T. denticola; abscess and F. alocis, T. forsythia, and T. denticola (all p < 0.05). The findings of this study indicated that F. alocis, T. forsythia, and T. denticola seem to be associated with endodontic signs and symptoms. Additionally, F. alocis and T. forsythia were detected more frequently in teeth with necrotic pulp than in teeth with failing endodontic treatment.
Contemporary Clinical Dentistry, 2016
Background: The detection frequency of Porphyromonas gingivalis and Treponema denticola in chronic periodontitis (CP) and aggressive periodontitis (AgP) is not explored well in Indian population. Aim: The study was undertaken to detect P. gingivalis and T. denticola in CP as well as in AgP patients using polymerase chain reaction (PCR), and to determine the relationship between the frequency of these two microorganisms and the severity of clinical periodontal parameters. Materials and Methods: Subgingival plaque samples were collected from ninety participants (thirty CP patients, thirty AgP patients, and thirty healthy participants) and the aforementioned two microorganisms were detected using PCR. Results: However, when CP and AgP were compared for the detection frequency of two microorganisms, no statistically significant difference was noted. A statistically significant increase in the number of bacteria-positive sites increased as the score of plaque index (PI), gingival index (GI), and clinical attachment level of CP and AgP patients increased. Coexistence of P. gingivalis and T. denticola was frequently observed in deep periodontal pockets. Conclusions: Study findings suggest that P. gingivalis and T. denticola are significantly associated with the severity of periodontal tissue destruction. Statistically significant association exists between clinical periodontal parameters such as PI, GI, periodontal pocket depth (PPD), and clinical attachment loss and presence of both the microorganisms.
Evaluation of role of periodontal pathogens in endodontic periodontal diseases
Journal of Family Medicine and Primary Care, 2020
Aim: This study aimed to correlate periodontal pathogens in endodontic periodontal diseases. Methodology: This study was conducted on 40 patients of both genders. All the participants were obtained from department of endodontics and periodontology with history of endo‑perio lesion in same teeth. Polymerase chain reaction was performed and correlation was established. Results: This study included 18 males and 22 females. The mean age of male was 42.5 years and female was 41.3 years. Specimens of Tannerella forsythia were isolated from 94% endodontium and 92% periodontium, Porphyromonas gingivalis from 71% endodontium and 55% periodontium, Aggregatibacter actinomycetemcomitans from 12% endodontium and 58% periodontium. The difference was significant (P < 0.05). Bacteria in endodontic‑periodontal infection confirmed statistically significant correlation between absolute quantitation of T. forsythia and P. gingivalis (r = 0.412, P < 0.05), P. gingivalis and A. actinomycetemcomitans (r = 0.524, P < 0.05), and T. forsythia and A. actinomycetemcomitans (r = 0.427, P < 0.05). Conclusion: There was correlation between targeted bacterial species levels from concurrent endodontic‑periodontal diseases. Thus, it can be suggested that dentinal tubules may be the pathway for spread of bacteria.
Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 2008
The aim of the present study was to evaluate the presence of the selected pathogens in samples from deciduous and permanent tooth root canals by using PCR method and to determine the association of these organisms with clinical symptoms. Study design. A total of 145 children, 5 to 13 years old, were involved in this study. The presence of selected pathogens (Actinomyces israelii, Candida albicans, Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Streptococcus intermedius, Treponema denticola, Parvimonas micra, Tannerella forsythensis, Enterococcus faecium, Prevotella melaninogenica) in infected root canals was studied using PCR. Results. T. denticola (P ϭ .012, .02) and E. faecalis (P ϭ .012, .04) were highly associated with periapical radiolucency and previous pain, while P. gingivalis was associated with tenderness to percussion in both deciduous and permanent teeth (P ϭ .01, .015). Conclusion. The results of the present study confirm that certain species of microorganisms are associated with clinical signs and symptoms of endodontic disease in both deciduous and permanent teeth.
Microorganisms involved in endodontic infection of permanent teeth: A systematic review
African Journal of Microbiology Research, 2013
This study investigated the microbial species present in necrotic pulps of permanent teeth needing endodontic treatment. The search for articles was conducted on the Health Science Database and considered all articles published until October 2011, with selected human clinical studies examined, through molecular biology the microorganisms that are present in root canals of permanent teeth requiring root canal treatment with necrosis pulps (exposition). The selected articles were categorized according to the methodological quality and evidence in levels A (high), B (moderate), and C (low). The search strategy was based on PubMed, Bireme, Cochrane and OVID databases. The data extracted from the studies were also tabulated. Eighty-four titles and abstracts were assessed and eight articles met the inclusion criteria, showing that there is a high diversity of microorganisms involved in necrotic permanent teeth. The microorganisms found in these articles were: Porphyromonas gingivalis (27.8%), Porphyromonas endodontalis (42.6%), Prevotella intermedia (5.6%), Prevotella nigrescens (7.4%), Bacteroides forsythus (21%) and Enterococcus faecalis (>50%). This systematic review found high evidence of the polymicrobial nature of primary endodontic infections in necrotic permanent teeth, and also showed that there is a dominance of anaerobic bacteria.
The Bulletin of Tokyo Dental College, 2000
The relationship between the detection of Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola in subgingival plaque samples of periodontal pockets and periodontal status was evaluated using the polymerase chain reaction (PCR). A total of 165 sites in 60 periodontitis patients were examined, and the relationships between the detection of each of the three bacterial species and the pocket depth and bleeding on probing (BOP) were analyzed. The detection ratios of P. gingivalis, B. forsythus, and T. denticola in samples from adult periodontitis lesions were 75.5%, 69.8%, and 72.6%, respectively. It was found that all sites where all three microorganisms were detected were BOP positive and had greater pocket depths than those where only one or two species were found. The detection rate of B. forsythus and T. denticola decreased with age in the sites in which PD was less than 4 mm. The present study indicates that detection of a mixed infection by P. gingivalis, B. for...
Journal of Dental Research, Dental Clinics, Dental Prospects, 2016
Background. The most common species isolated from primary endodontic infections are black-pigmented bacteria. These species are implicated in apical abscess formation due to their proteolytic activity and are fastidious in nature. Therefore, the present study was carried out to evaluate the presence and identification of various pigmented Porphyromonas and Prevotella species in the infected root canal through culture-based techniques. Methods. Thirty-one patients with primary endodontic infections were selected. Using sterile paper points, samples were collected from the root canals after access opening and prior to obturation, which were cultured using blood and kanamycin blood agar. Subsequently, biochemical test was used to identify the species and the results were analyzed using percentage comparison analysis, McNemar and chi-squared tests, Wilcoxon match pair test and paired t-test. Results. Out of 31 samples 26 were positive for black-pigmented organisms; the predominantly isolated species were Prevotella followed by Porphyromonas. In Porphyromonas only P. gingivalis was isolated. One of the interesting features was isolation of P. gingivalis through culture, which is otherwise very difficult to isolate through culture. Conclusion. The presence of Prevotella and Porphyromonas species suggests that a significant role is played by these organisms in the pathogenesis of endodontic infections.