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。     

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。     

Microorganisms in root canal-treated teeth from a German population

Posttreatment apical periodontitis is usually associated with persistent or secondary intraradicular infection. This study evaluated the presence and relative levels of 28 bacterial taxa in treated root canals of teeth with posttreatment apical periodontitis from German patients using 16S ribosomal RNA (rRNA) gene probes in a reverse-capture checkerboard hybridization assay. Species-specific polymerase chain reaction (PCR) was also performed to detect Enterococcus faecalis and Candida albicans. Bacterial DNA was detected in all samples. Twenty of the 28 taxon-specific probes tested were reactive with at least one sample. Taxa detected more frequently included Streptococcus species (47%), Lactobacillus species (35%), Dialister invisus (29%), Eubacterium infirmum (29%), Prevotella intermedia (29%), Selenomonas sputigena (29%), Synergistes oral clone BA121 (29%), and Treponema denticola (29%). Only eight taxa were present at levels >10(5). Of these, streptococci and T. denticola were the most prevalent. Species-specific PCR detected E. faecalis in 47% of the cases and C. albicans in 6%. Findings of this study confirm the strong association between persistent/secondary intraradicular infection and posttreatment apical periodontitis. Most cases harbored a mixed infection, and E. faecalis, if present, was never the most dominant species in the consortium. Several other bacterial taxa were detected, and an involvement with the etiology of posttreatment apical periodontitis is suspected.     

Microbial analysis of canals of root-filled teeth with periapical lesions using polymerase chain reaction

The objective of the present study was to investigate the presence of nine bacterial species in root-filled teeth associated with periapical lesions using a polymerase chain reaction analysis and to correlate these species with clinical features of the cases. DNA was extracted from 45 canal samples of root-filled teeth with periapical lesions. A PCR assay using species-specific primers of 16S rDNA and the downstream intergenic spacer region was used for microbial detection. Enterococcus faecalis was the most prevalent species, detected in 77.8% of the study teeth, followed by Peptostreptococcus micros, detected in 51.1%. Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, and Prevotella nigrescens were detected in 35.6%, 22.2%, 11.1%, and 11.1% of the sampled teeth, respectively. Moreover, PCR detected Filifactor alocis in 26.7%, Treponema denticola in 24.4%, and Tannerella forsythia in 4.4% of the samples. T. denticola and P. micros were statistically associated with tenderness to percussion (p < 0.05). P. nigrescens was associated with the presence of spontaneous pain and abscess (p < 0.05). P. endodontalis and P. nigrescens were associated with purulent exudates (p < 0.05). Synergistic relationship was also observed between some species. The results of this study indicated that E. faecalis was the most frequently identified test species by PCR in teeth with failing endodontic treatment.     

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.     

The presence of periodontopathogens associated with the tumour necrosis factor-alpha expression in patients with different periodontal status

The purpose of this study was to investigate the relationship between P gingivalis, T forsythia, T denticola, P intermedia, and A. actinomycetemcomitans in the sulci or pockets of patients with gingivitis (G), mild chronic periodontitis (MiCP), moderate chronic periodontitis (MoCP) and severe periodontitis (SP), and the expression of TNF-alpha in gingival tissue associated with clinical parameters. Six patients with G, 7 with MiCP, 23 with MoCP and 7 with SP were recruited. Pathogens obtained from the sulci or pockets were identified by PCR, and expression of TNF-alpha from gingival tissue was analysed Probing depth (PD), clinical attachment level (CAL) and loss of bone were recorded. P gingivalis was detected at the following rates: 16.6% in subjects with G 57.1% in MiCP 57.8 % in MoCP and 58.1% in SP (p < 0.05). P intermedia was not identified in subjects with G A. actinomycetemcomitans was only identified in subjects with MoCP (31.5%) and SP (42.8%). T denticola and T forsythia were identified in all subject groups. Bacterial combinations were identified as follows: P denticola + P intermedia and PR intermedia + T forsythia were associated (p = 0.04, p = 0.02) with the presence of TNF-alpha mRNA in 20% and 25% of subjects, respectively. P gingivalis + A. Actinomycetemcomitans andA. actinomycetemcomitans + T forsythia were associated with severe PD and CAL, respectively. The association between the presence of P intermedia and expression levels of TNF-alpha was significant (p = 0.05). These results indicate that the proportion of patients with P gingivalis increases with the progression of disease. We observed that the presence of P intermedia may trigger the expression of TNF-alpha and cause a worsening of the patient's clinical status.     

根管治疗期间急性发作与慢性根尖周炎患牙根管细菌组成比较

目的:比较根管治疗期间炎症急性发作及慢性根尖周炎患牙根管内8种厌氧菌检出情况,分析急性发作时根管内定植细菌种类及其相关性。方法:分别提取26例根管预备后约诊期间炎症急性发作患牙根管样本和23例慢性根尖周炎患牙根管样本,提取样本细菌DNA,利用细菌16S rRNA引物通过PCR扩增方法鉴定细菌。结果:慢性根尖周炎样本根管细菌检出率达100%(23/23),根管治疗期间急性发作样本细菌检出率为92.31%(24/26);产黑普氏菌、齿垢密螺旋体和直肠弯曲杆菌在急性发作样本中的检出率较慢性根尖周炎样本显著增高(P<0.05)。急性发作样本中牙龈卟啉菌与福赛氏类杆菌的检出显著相关(OR>2,P<0.05)。结论:根管内感染是根管治疗期间炎症急性发作的重要原因,厌氧菌在急性发作的发生中起重要作用。     

Identification of oral spirochetes at the species level and their association with other bacteria in endodontic infections

OBJECTIVES: Recent molecular approaches have revealed that fastidious organisms such as Bacteroides forsythus and oral treponemes were frequently found in root canals with apical periodontitis. The purpose of this study was to identify the isolates of oral spirochetes at the species level in endodontic infections and to determine their association with B forsythus and Porphyromonas gingivalis. STUDY DESIGN: Seventy-nine teeth with apical periodontitis were selected for this study. After sampling from the root canals aseptically, polymerase chain reaction amplification for the 16S rRNA gene was performed with eubacterial universal primers. Subsequently, dot-blot hybridization was performed with 8 species-specific oligonucleotide probes. The microbial associations were analyzed by using the odds ratio. RESULTS: The most frequently found species was P gingivalis (27.4%), followed by Treponema maltophilum (26%), B forsythus (16.4%), and Treponema socranskii (2.7%). Other treponemes, including Treponema denticola, were not detected in our samples. Significant microbial associations were identified between T maltophilum, B forsythus, and P gingivalis by performing analysis with the odds ratio. CONCLUSION: Our results indicate that T maltophilum should be included in etiologic studies of endodontic diseases.     

The distribution of periodontopathic bacteria among Japanese children and their parents

OBJECTIVE AND BACKGROUND: It is not well known how periodontopathic bacteria colonize in the oral cavity during childhood. The purpose of this study was to investigate the distribution of periodontopathic bacteria in oral cavities of children and their parents and the relationship between the bacterial findings and clinical parameters. METHODS: Fifty-six children (mean age: 8.3 +/- 3.5, range: 1-15 years), including 15 with deciduous dentition, 26 with mixed dentition and 15 with permanent dentition, and their parents participated in this study. Whole saliva and dental plaque of the children and whole saliva of their parents were collected for detection of seven species of periodontopathic bacteria (Actinobacillus actinomycetemcomitans, Tannerella forsythensis (Bacteroides forsythus), Campylobacter rectus, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Treponema denticola) using the polymerase chain reaction method. Clinical parameters including simplified Oral Hygiene Index and Papillary-Marginal-Attachment Index were recorded for the children and their accompanied parents. RESULTS: The detection frequencies of T. forsythensis, C. rectus, P. nigrescens, T. denticola, A. actinomycetemcomitans and P. gingivalis in the oral cavities of children were 42.9%, 94.6%, 42.9%, 48.2%, 1.8% and 8.9%, respectively. T. forsythensis, P. gingivalis and T. denticola were detected more frequently in the saliva of parents (54.8%, 54.8%, 88.1%, respectively) than in the saliva of children (25.5%, 7.3%, 41.8%, respectively). Different detection frequencies of P. nigrescens were found among the oral cavities of children with deciduous, mixed and permanent dentitions. In mixed dentition, females harbored T. forsythensis more frequently than males did. Children who harbored T. forsythensis, P. intermedia, P. nigrescens and T. denticola showed high scores for oral debris measurement by simplified Oral Hygiene Index. T. forsythensis, P. intermedia and P. nigrescens were detected more frequently in children whose parents were positive for these pathogens than in children whose parents were negative. CONCLUSIONS: High plaque retention seems to promote the colonization of periodontal pathogens in the oral cavities of children. T. forsythensis, P. intermedia and P. nigrescens were detected more frequently in the oral cavities of children whose parents already harbored these bacteria. Familial transmission of these bacteria is suggested.     

Association of black-pigmented bacteria with endodontic infections.

Black-pigmented bacteria (BPB) have been associated with endodontic infections. The purpose of this study was to evaluate further the presence of BPB with the clinical signs and symptoms associated with endodontic infections. Microbial samples were collected from the root canals of 40 intact teeth with necrotic pulps and apical periodontitis. Conventional laboratory methods were used for identification of the strains of BPB isolated in pure culture. In addition, the polymerase chain reaction and specific primers for 16S r-RNA genes were used to differentiate Prevotella nigrescens from Prevotella intermedia. Twenty-two (55%) samples were positive for the growth of BPB. Of those, 11 of 22 (50%) were identified as P. nigrescens, 8 of 22 (36%) were P. intermedia, 2 of 22 (9%) were Porphyromonas gingivalis, and 1 of 22 (5%) was Prevotella melaninogenica. Sixteen of the 22 root canals positive for the growth of BPB were associated with purulent drainage either from the root canal or an associated sinus tract. Statistical analysis did not show a significant relationship for the presence of BPB with clinical signs and symptoms.     

Selected endodontic pathogens in the apical third of infected root canals: a molecular investigation

Bacteria located at the apical portion of the root canals are conceivably in a strategic position to induce damage to the periradicular tissues and resulting inflammatory diseases. This study sought to investigate the prevalence of 11 selected putative endodontic pathogens in the apical third of infected root canals associated with periradicular lesions. The apical root portion of 23 extracted teeth with carious pulpal exposures and attached periradicular lesions was sectioned, and the root canals were sampled for microbiological investigation. DNA was extracted from the samples and analyzed for the presence of 11 bacterial species using a nested polymerase chain reaction assay. The results showed that Pseuramibacter alactolyticus occurred in 10 cases (44%), Treponema denticola in 6 (26%), Fusobacterium nucleatum in 6 (26%), Porphyromonas endodontalis in 4 (17%), Filifactor alocis in 2 (9%), Dialister pneumosintes in 1 (4%), Porphyromonas gingivalis in 1 (4%), and Tannerella forsythensis in 1 (4%). No sample yielded Prevotella intermedia, Prevotella nigrescens, or Campylobacter rectus. Of the samples examined, 17 were positive for at least 1 of the target species. Occurrence of these bacterial species in the apical third of infected root canals suggests that they can be involved in causation of periradicular lesions.     

Comparison of profiles of key periodontal pathogens in periodontium and endodontium

Abstract – Despite the established anatomical relationship between the periodontal and pulpal tissues, bacterial migration between endodontium and periodontium is still under discussion. The objective of this study was an investigation of profiles of periodontal pathogens in pulpal and periodontal diseases affecting the same tooth by means of 16S rRNA gene directed polymerase chain reaction (PCR). 31 intact teeth with both pulp and marginal infections were investigated. The diagnosis was based on clinical and radiological examination. Samples were taken from the gingival sulcus or periodontal pocket, respectively, with sterile paper points before trepanation of the teeth. After trepanation sterile paper points and Hedstroem files were used for taking samples from the root canal. Specific PCR methods were used to detect the presence of the following pathogens: Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Treponema denticola. In addition, quantitative competitive PCR was used to determine the total bacterial count of the samples. The investigated pathogens were proven to be present in the endondontium in all disease categories. Particularly in endodontic samples of "chronic apical periodontitis" and "chronic adult periodontitis" profiles of the periodontal pathogens were found. The results confirmed that periodontal pathogens often accompany endodontic infections and supported the idea that the periodontic-endodontic interrelationships should be considered as critical pathways which might contribute to refractory courses of endodontic or periodontal diseases.     

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.     

Comparison of profiles of key periodontal pathogens in periodontium and endodontium

Despite the established anatomical relationship between the periodontal and pulpal tissues, bacterial migration between endodontium and periodontium is still under discussion. The objective of this study was an investigation of profiles of periodontal pathogens in pulpal and periodontal diseases affecting the same tooth by means of 16S rRNA gene directed polymerase chain reaction (PCR). 31 intact teeth with both pulp and marginal infections were investigated. The diagnosis was based on clinical and radiological examination. Samples were taken from the gingival sulcus or periodontal pocket, respectively, with sterile paper points before trepanation of the teeth. After trepanation sterile paper points and Hedstroem files were used for taking samples from the root canal. Specific PCR methods were used to detect the presence of the following pathogens: Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Treponema denticola. In addition, quantitative competitive PCR was used to determine the total bacterial count of the samples. The investigated pathogens were proven to be present in the endondontium in all disease categories. Particularly in endodontic samples of "chronic apical periodontitis" and "chronic adult periodontitis" profiles of the periodontal pathogens were found. The results confirmed that periodontal pathogens often accompany endodontic infections and supported the idea that the periodontic-endodontic interrelationships should be considered as critical pathways which might contribute to refractory courses of endodontic or periodontal diseases.     

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

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

Polymerase chain reaction of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in primary endodontic infections

The aim of this study was to investigate the correlation between endodontic clinical signs and symptoms and the presence of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia or their association by nested polymerase chain reaction assay. Microbial samples were taken from 50 cases with necrotic pulp tissues in primary infections. DNA was extracted from the samples, which were analyzed for the presence of three endodontic pathogens by using species-specific primers. P gingivalis, T denticola, and T forsythia were detected in 46%, 38%, and 22% of the symptomatic cases, respectively. The bacterial complex composed by T forsythia, P gingivalis, and T denticola was found in 14% of the cases with spontaneous pain, tenderness to percussion, swelling, and pain on palpation. The high prevalence of P gingivalis, T denticola, and T forsythia in the samples examined suggests that these bacteria are related to the etiology of symptomatic periradicular diseases.     

再治疗根管粪肠球菌毒力因子gelE表达与临床表现的关系研究

目的:比较再治疗根管内粪肠球菌(enterococcus faecalis)毒力因子gelE表达情况,分析gelE表达与临床表现的关系。方法:采集临床需要根管再治疗病例的根管内细菌样本53例,利用Real Time Quantitative PCR技术来检测gelE的表达情况。统计学分析gelE表达与患者临床表现之间的关系。结果:再治疗根管内粪肠球菌毒力因子gelE在有临床症状或体征和有根尖暗影的病例中表达增强(P<0.05)。结论:再治疗根管内粪肠球菌毒力因子gelE的表达增强与临床症状或体征的出现有关系。     

Prevalence and clonal analysis of Porphyromonas gingivalis in primary endodontic infections

This study investigated the prevalence of Porphyromonas gingivalis in 62 teeth with primary endodontic infections by using a species-specific 16S rRNA gene-based nested polymerase chain reaction assay. P. gingivalis isolates recovered from 2 infected root canals were also analyzed for clonal diversity by using arbitrarily primed PCR. Overall, P. gingivalis was found in 48% of the samples. This species was specifically detected in 36% of canals of teeth with chronic apical periodontitis, in 46% of the cases of acute apical periodontitis, and in 67% of acute apical abscesses. P. gingivalis was significantly more frequent in abscess aspirates than in canals of teeth with chronic apical periodontitis (P < .05). Typing of colonies retrieved from 2 infected canals revealed 2 clones per individual. These findings confirmed that P. gingivalis can be an important endodontic pathogen, mostly associated with acute abscesses, and demonstrated that different clonal types of this species can colonize the root canal in the same individual.     

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.     

Endodontic management of immature teeth with spontaneous apical closure and periapical lesions: case series and review of the literature

Spontaneous apical closure in non‐vital immature teeth has been rarely encountered and outcome of non‐surgical endodontic treatment of related teeth associated with periapical lesions has not yet been adequately elucidated. The aim of this article was to report endodontic management of spontaneous apical closure of infected untreated immature teeth with periapical lesions and to review previously proposed mechanisms for the development of spontaneous hard tissue barrier. Three patients were referred at different time intervals to the endodontic clinic for treatment of their maxillary anterior incisors with acute or chronic apical periodontitis . Dental histories indicated that related teeth had been subjected to trauma approximately 12–18 years previously. Radiographically, the involved teeth exhibited incomplete root formation with spontaneous apical closure and were associated with an apical radiolucency. After biomechanical preparation, calcium hydroxide paste was applied and was changed once or twice within 3 months. All canals were then filled with gutta‐percha and AH Plus and the follow‐up period was 16–50 months; both clinical and radiographic examinations revealed adequate function, the absence of clinical symptoms and significant healing of the periapical radiolucency.