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。     

牙周基础治疗对慢性牙周炎患者临床指标及5种牙周可疑致病微生物的影响

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

不同牙周状况龈下菌斑中牙龈卟啉单胞菌和福赛斯坦纳菌的分布

目的 分析中国牙周健康者和慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌和福赛斯坦纳菌的分布情况,探讨两种细菌在中国慢性牙周炎患者中的分布及其致病机理.方法 收集65例牙周健康者、62例慢性牙周炎患者的龈下菌斑,采用16S rRNA PCR方法检测牙龈卟啉单胞菌和福赛斯坦纳菌的分布.结果 牙周健康者牙龈卟啉单胞菌和福赛斯坦纳菌的检出率分别是21.6%、12.3%,慢性牙周炎患者病变位点两菌检出率分别为74.2%、58.1%,牙周炎健康位点两菌的检出率分别是22.6%、4.8%.牙周炎病变位点两种细菌的检出率均明显高于牙周健康者和牙周炎健康位点(P＜0.001);福赛斯坦纳菌在牙周健康者中的检出率高于牙周炎健康位点(P＜0.05);牙周健康者和牙周炎健康位点牙龈卟啉单胞菌的检出率没有差异.慢性牙周炎患者两菌联合检出率为51.6%.结论 牙龈卟啉单胞菌、福赛斯坦纳菌以及两种细菌联合感染与牙周炎密切相关.     

纸尖法和刮匙法对龈下牙周致病菌检出比较研究

目的：比较纸尖法和刮匙法对慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌、福赛斯坦纳菌、伴放线菌嗜血菌、具核梭杆菌检出的影响。方法：对30例慢性牙周炎患者分别用纸尖和刮匙法收集4个区域牙周袋探诊最深位点龈下菌斑，采用16SrRNAPCR方法检测两种方法收集到的龈下菌斑样本中的牙龈卟啉单胞菌、福赛斯坦纳菌、伴放线菌嗜血菌、具核梭杆菌。结果：两种方法一致性较好，纸尖法与刮匙法无显著差异（P〉0．05）。结论：纸尖法与刮匙法均为比较准确的取样方法。     

不同采样方法对龈下牙周致病菌检出的比较研究

目的:比较4-site法和full-mouth法对慢性牙周炎病人龈下菌斑中牙龈卟啉单胞菌、福赛斯坦纳菌、伴放线菌嗜血菌、具核梭杆菌检出的影响.方法:对30例慢性牙周炎病人用纸尖收集4个区牙周袋探诊最深位点龈下菌斑(4-site法)和收集全口龈下菌斑(full-mouth法),采用16S rRNA PCR方法检测两种方法收集到的龈下菌斑样本中的牙龈卟啉单胞菌、福赛斯坦纳菌、伴放线菌嗜血菌、具核梭杆菌.结果:两种方法一致性较好,4-site法与full-mouth法无显著差异(P>0.05).结论:4-site法采集到的龈下菌斑具有良好的代表性.     

慢性牙周炎患者牙周袋内硫化物水平与厌氧细菌分布的相关性研究

目的:探讨慢性牙周炎患者牙周袋内硫化物水平(sulfide levels in periodontal pockets,SUL)与牙龈卟啉单胞菌、中间普氏菌、伴放线放线杆菌分布的相关性.方法:用perio2000 system金刚探针牙周诊断仪测定慢性牙周炎患者SUL,采用PCR方法检测龈下菌斑中牙龈卟啉单胞菌、中间普氏菌和伴放线放线杆菌.结果:慢性牙周炎患者随着牙周袋内SUL浓度的增加牙龈卟啉单胞菌、中间普氏菌的检出率逐渐增加,相关系数分别为0.812和0.651(P<0.05);SUL阳性位点与阴性位点中牙龈卟啉单胞菌的检出率分别是为85.6%、26.7%,中间普氏菌的检出率分别为95.2%、53.3%.SUL阳性位点中牙龈卟啉单胞菌和中间普氏菌的检出率明显高于SUL阴性位点;在80.9%的SUL阳性位点中牙龈卟啉单胞菌与中间普氏菌共存.SUL阳性位点与阴性位点均未检出伴放线放线杆菌.结论:慢性牙周炎患者SUL与牙龈卟啉单胞菌和中间普氏菌分布关系较为密切.     

3种寡核苷酸探针对龈下菌斑中牙周致病菌的检测

目的　采用寡核苷酸探针研究龈下菌斑中3种牙周致病菌的分布。方法　利用3种寡核苷酸探针对 60例慢性牙周炎患者60个患病位点、10例健康人的10个健康对照位点龈下菌斑中的牙龈卟啉单胞菌、福赛类杆菌、牙密螺旋体进行检测。结果　牙周炎位点龈下菌斑中的牙龈卟啉单胞菌、福赛类杆菌、牙密螺旋体的检出率分别为91·67%,90·00%和95·67%,明显高于健康对照位点;有83·33%的牙周炎位点同时检出3种致病菌,3种细菌检出情况为两两正相关(P<0·01)。结论　牙龈卟啉单胞菌、福赛类杆菌、牙密螺旋体在慢性牙周炎患者龈下菌斑中的检出率很高,它们间可能存在相互协同致病作用。     

Er: YAG激光对2型糖尿病相关性牙周炎患者龈下菌群动态变化的研究

目的 动态观察2型糖尿病相关性牙周炎患者经Er: YAG激光治疗前后龈下菌群的变化,并与不伴全身疾病的慢性牙周炎龈下菌群进行比较。方法 收集11例2型糖尿病相关性牙周炎患者的13对患牙（26个位点）作为试验组,分别进行Er: YAG激光治疗和超声治疗,采集治疗前及治疗后1、3个月的龈下菌斑;同时收集11例牙周状况相近的不伴全身疾病的中重度慢性牙周炎患者13个位点的龈下菌斑作为对照组,分析试验组是否有菌群种类变化。提取龈下菌斑DNA,进行变性梯度凝胶电泳分离及条带回收测序。结果 试验组与对照组患牙龈下菌斑的优势致病菌存在差异,分别为中间普雷沃菌和福赛斯坦纳菌。激光组治疗前后龈下菌群构成也发生改变,治疗后1个月,有的条带表达减弱或消失,并有新条带出现;测序结果表明,新出现的条带为放线菌的一种,减弱、消失的为福赛斯坦纳菌。结论 Er: YAG激光治疗前后龈下菌群的构成发生变化,治疗后1个月是关键时期;治疗后3个月,激光治疗在阻止细菌再定植方面可能更具优势。     

不同牙周状况龈下菌斑中齿垢密螺旋体的分布

目的：分析慢性牙周炎、牙龈炎患者和牙周健康者龈下菌斑中齿垢密螺旋体的分布情况,探讨该微生物与不同牙周状况的关系。方法：收集12例慢性牙周炎、5例牙龈炎患者和5例牙周健康者,共70个位点的龈下菌斑,采用Taq Man16SrRNA实时荧光定量PCR技术检测齿垢密螺旋体的分布。结果：慢性牙周炎病变位点齿垢密螺旋体检出率是86％,牙龈炎的是86％,牙周健康的是1.0％,3组检出率之间的差异没有统计学意义;微生物的检出量经对数转换后,3组之间及两两比较均有统计学意义。结论：TaqMan实时荧光定量PCR技术检测微生物灵敏度高,检出率高;龈下菌斑中齿垢密螺旋体与牙周状态密切相关。     

牙周炎的厌氧菌群的分离及对抗生素的敏感性研究

目的：调查牙周炎思考口腔厌氧菌群的分布并研究其对β内酰胺药物的敏感性。方法：选择44例牙周炎患者，对其龈下菌斑中的厌氧菌进行分离培养，用7种β内酰胺药物对分离到的厌氧菌进行敏感性试验。结果：与牙周病有关的厌氧菌分离率为100％，产黑色素普雷沃菌86．4％，中间普雷沃菌56．8％，具核梭杆菌63．6％，放线菌38．6％，微小消化链球菌52．3％，牙龈卟啉单胞菌65．9％，二氧化碳嗜纤维菌9．1％。厌氧菌对各类β内酰胺药物保持敏感。结论：产黑色素普雷沃菌、中间普雷沃菌、具核梭杆菌、微小消化链球菌、牙龈卟啉单胞菌和放线菌，是牙周炎思考牙龈沟中的优势菌；β内酰胺药物可作为厌氧菌牙周炎的治疗药物。     

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.     

Description of the subgingival microbiota of periodontally untreated Mexican subjects: chronic periodontitis and periodontal health

BACKGROUND: Recent studies have suggested that changes in the prevalence and/or proportion of distinct microorganisms characterize the subgingival microbial profiles of populations around the world. At present, no information is available on the subgingival microbiota of Mexican subjects. The purpose of the present study was to determine the microbial composition of subgingival plaque in Mexican subjects with untreated chronic periodontitis. METHODS: A total of 44 chronic periodontitis and 20 periodontally healthy subjects (who were currently non-smokers) were selected. Clinical measurements including plaque accumulation, gingival erythema, bleeding on probing, suppuration, probing depth, and attachment level were recorded at six sites of every tooth. Up to 28 subgingival plaque samples were obtained from each subject and individually analyzed to determine the levels, proportion, and prevalence of 40 microbial species using the checkerboard DNA-DNA hybridization technique. RESULTS: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythensis were the only species that presented higher mean levels in periodontitis subjects. The proportions of P. gingivalis (P<0.001), T. forsythensis (P<0.01), and red complex species (P. gingivalis, T. forsythensis, and T. denticola; P<0.001) as a group were also significantly higher in periodontitis subjects. Periodontally healthy subjects harbored a significantly larger proportion of Actinomyces species (P<0.05). No significant differences were detected in the percentage of carriers of any of the species tested. CONCLUSIONS: Our results revealed that the subgingival microbiota of untreated chronic periodontitis Mexican subjects was characterized by increases in the level, prevalence, and proportion of classic periodontal pathogens. However, the prevalence and proportion of specific microbial species varied significantly from the results of other reports on subjects from different geographical locations.     

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.     

Clustering of subgingival microbial species in adolescents with periodontitis

It has become increasingly recognized that groups of microorganisms interact within the subgingival plaque of adult subjects with periodontitis. It is much less clear, however, whether the consortia of microorganisms associated with periodontitis are different in early and more advanced cases of periodontitis. To investigate this point further, subgingival plaque was collected from six sites in 87 adolescents with periodontitis and 73 controls and the samples were analyzed for the detection of 18 microbial species using the DNA-DNA hybridization technique. Actinomyces oris accounted for the highest proportion of the flora and was more predominant among controls. Prevotella nigrescens, Prevotella intermedia, Porphyromonas gingivalis, and Tannerella forsythia were present at higher levels among the subjects with periodontitis. Factor analyses identified one factor characterized by highly positive loadings for T. forsythia, Campylobacter rectus, P. gingivalis, P. intermedia, P. nigrescens, Parvimonas micra, and Treponema denticola, and another factor characterized by highly positive loadings of A. oris, Capnocytophaga ochracea, Eikenella corrodens, Streptococcus intermedius, Selenomonas noxia, Streptococcus oralis, Streptococcus sanguinis, and Veillonella parvula. Aggregatibacter actinomycetemcomitans and Streptococcus mutans did not load on any of the two factors, while Fusobacterium nucleatum loaded on both. These findings confirm the occurrence of clustering of subgingival bacteria according to case status also among young individuals.     

Quantitative analysis of periodontal pathogens in aggressive periodontitis patients in a Japanese population

The aim of the present study was to clarify the relationship between the relative/absolute numbers of periodontal bacteria and different types of periodontitis. Fifteen patients with localized aggressive periodontitis (LAgP), 25 patients with generalized aggressive periodontitis (GAgP) and 28 patients with chronic periodontitis (CP) were included in this study. Saliva and subgingival plaque samples were collected from all subjects for microbiological analysis. The prevalence and proportions of Actinobacillus actinomycetemcomitans, Tannerella forsythensis, Porphyromonas gingivalis and Treponema denticola were determined by conventional PCR and real-time PCR. The prevalence of A. actinomycetemcomitans in saliva was significantly higher in LAgP patients (46.7%) and GAgP patients (40.0%) than that in CP patients (14.3%). The mean proportion of A. actinomycetemcomitans in LAgP patients (4.42%) was significantly higher than that in GAgP patients (0.59%) and CP patients (0.37%) in saliva. In subgingival plaque, LAgP patients showed a significantly higher mean proportion of T. forsythensis (19.8%) than CP patients (7.45%). In conclusion, A. actinomycetemcomitans was the more predominant periodontopathic bacteria in LAgP than in GAgP and CP. The increased proportion of T. forsythensis might relate to LAgP, in addition to A. actinomycetemcomitans. These results indicate that real-time PCR analysis is useful for the evaluation of the bacterial profiles in different types of periodontitis.     

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.     

Treponema socranskii, Treponema denticola, and Porphyromonas gingivalis are associated with severity of periodontal tissue destruction.

BACKGROUND: The aim of the present study was to identify Treponema socranskii in addition to Treponema denticola and Porphyromonas gingivalis by polymerase chain reaction (PCR), and to clarify the relationship between the presence of these microorganisms and the severity of clinical periodontal parameters. METHODS: Saliva and subgingival plaque collected from 123 subjects (38 aggressive periodontitis patients, 65 chronic periodontitis patients, 20 healthy patients) were subjected to PCR to detect the aforementioned 3 microorganisms. RESULTS: Detection frequencies of T. socranskii, T. denticola, and P. gingivalis in plaque samples from aggressive periodontitis patients (71.1%, 73.7%, 84.2%, respectively) and chronic periodontitis patients (89.2%, 93.8%, 95.3%) were much higher than those from healthy subjects (30%, 5.0%, 10.0%). In aggressive and chronic periodontitis patients, these 3 species of bacteria were detected frequently at sites that showed deep periodontal pockets and severe attachment loss. The percentage of these bacteria-positive sites increased as the gingival index score of chronic periodontitis patients increased. T. socranskii was frequently detected together with T. denticola or P. gingivalis at the same sites, and coexistence of these microorganisms was frequently observed in deep periodontal pockets of aggressive periodontitis patients. CONCLUSIONS: T. socranskii, T. denticola, and P. gingivalis were frequently detected in periodontitis patients by PCR. The prevalence of these 3 microorganisms was correlated with various clinical parameters. Taken together, our findings suggest that T. socranskii, T. denticola, and P. gingivalis are associated with the severity of periodontal tissue destruction.     

Demographic, clinical, and microbial aspects of chronic and aggressive periodontitis in Colombia: a multicenter study

BACKGROUND: The microbial profile of periodontal disease varies among different human populations. This study evaluated the demographic, clinical, and microbiologic aspects of periodontitis in a multigeographic sample in Colombia. METHODS: Three hundred twenty-five patients with chronic periodontitis (CP), 158 patients with aggressive periodontitis (AgP), and 137 healthy-gingivitis controls from five regions of the country were studied. Clinical, microbial, and sociodemographic data were collected. Microbiologic identification was performed using polymerase chain reaction 16S rRNA gene on pooled subgingival samples, and the presence of Gram-negative enteric rods was evaluated by culture. Bivariate and multivariate logistic regression analyses were conducted. RESULTS: Porphyromonas gingivalis occurred in 71.5% of individuals with periodontitis, Tannerella forsythensis occurred in 58.5%, Campylobacter rectus occurred in 57.5%, Actinobacillus actinomycetemcomitans occurred in 23.6%, and enteric rods occurred in 34.5%. P. gingivalis was more common in CP and AgP than controls. A. actinomycetemcomitans was increased in AgP compared to controls and patients with CP. T. forsythensis, C. rectus, and Eikenella corrodens had a low presence in the West Pacific and Central regions, and enteric rods were increased in the Central region (P <0.05). Other sociodemographic factors were not associated with these microorganisms. CONCLUSIONS: Geographic regions do not influence the microbiota, but the microbiota may vary by geographic region. P. gingivalis, T. forsythensis, and C. rectus are the most prevalent periodontophatic microorganisms in Colombia. A. actinomycetemcomitans was more common in AgP, and a large percentage of the population studied had enteric rods in the subgingival plaque.