慢性牙周炎龈下菌斑中福赛斯坦纳菌的检测

目的检测慢性牙周炎患者和牙周健康者龈下菌斑中福赛斯坦纳菌的数量和所占比例,探讨福赛斯坦纳菌与牙周炎发生发展的关系。方法采集经常规聚合酶链反应（PCR）法检测福赛斯坦纳菌为阳性的61例慢性牙周炎患者和12例牙周健康者的龈下菌斑,应用TaqMan实时荧光定量PCR法对菌斑的细菌总量和福赛斯坦纳菌的数量进行定量检测,构建含有福赛斯坦纳菌和真细菌靶基因的重组质粒,建立定量标准。结果本研究设计的引物和探针具有良好的特异性和敏感性;慢性牙周炎患者病变位点的福赛斯坦纳菌数量和细菌总量均高于牙周健康者的健康位点,且福赛斯坦纳菌在龈下菌斑中的比例也比健康位点高（P<0.05）;龈下菌斑的细菌数量与探诊深度呈正相关（P<0.001）;龈下菌斑中福赛斯坦纳菌所占比例在不同的探诊深度间无统计学差异（P>0.05）。结论龈下菌斑中福赛斯坦纳菌的数量与牙周状况有密切关系,实时荧光定量PCR法是研究牙周病病因及治疗方法的有效手段。     

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.     

慢性牙周炎患者龈下菌斑中福赛斯坦纳菌与临床指标的关系

目的分析慢性牙周炎患者龈下菌斑中福赛斯坦纳菌（T.forsythensis）的分布情况,及其与牙周临床指标的关系,探讨T.forsythensis与慢性牙周炎发生发展的关系。方法采集108例慢性牙周炎患者的216个病变位点和108个健康位点的龈下菌斑,共324个样本,采用16S rRNA聚合酶链反应检测T.forsythensis的分布情况。对324个取样位点进行临床检查,记录探诊深度、临床附着丧失和探诊出血情况,分析T.forsythensis与临床指标的相互关系。结果慢性牙周炎患者病变位点与健康位点T.forsythensis的检出率分别为59.72%和3.70%,二者间有统计学差异（P<0.001）。随着探诊深度和临床附着丧失的增加,T.forsythensis的检出率也升高,Spearman相关系数分别为0.48和0.51。探诊出血阳性位点T.forsythensis的检出率为61.31%,明显高于探诊出血阴性位点的检出率8.80%（P<0.001）。结论T.forsythensis与慢性牙周炎发生、发展关系密切。     

龈沟产线菌检出率与牙周健康状况的相关性分析

目的 分析不同牙周健康状况者龈下菌斑中龈沟产线菌的检出率差异及其与牙周临床指标间的相关关系。方法 应用聚合酶链反应法分别检测17例牙周健康者的68个牙周健康位点、19例菌斑性牙龈炎患者的64个牙周健康位点和76个病变位点、14例慢性牙周炎患者的36个牙周健康位点和56个病变位点的龈下菌斑中龈沟产线菌的检出率,并分析检出率与牙周临床检查指标之间的相关关系。结果 龈沟产线菌的检出率在不同人群的健康位点组及不同人群疾病位点组依次分别增高,其中牙周健康者龈下菌斑中龈沟产线菌检出率最低,为30.88%（21/68）,而慢性牙周炎患者病变位点组龈下菌斑中龈沟产线菌检出率最高,为91.07%（51/56）。龈沟产线菌与位点发生牙周病变之间的相关性有统计学意义（P<0.000 1）,其检出率与位点牙龈出血指数、临床探诊深度、临床附着水平的比值比分别为5.26、8.85、11.65。结论 牙周炎患者口腔中携带龈沟产线菌的风险升高;局部携带龈沟产线菌增加了位点牙周临床指标（牙龈出血指数、临床探诊深度、临床附着水平）异常的风险。     

福赛斯坦纳菌与牙周洁刮治疗效的关系

目的:分析福赛斯坦纳菌与牙周洁刮治术(SRP)疗效的关系。方法:采集20例慢性牙周炎患者(平均年龄44.33±13.86岁)SRP治疗前后口内4个区牙周袋最深位点的龈下菌斑,并记录临床指标,应用TaqMan实时荧光定量PCR法定量检测样本中的福赛斯坦纳菌。结果:SRP治疗能有效改善临床指标(P<0.001),降低福赛斯坦纳菌的检出率和数量水平(P<0.001);治疗后福赛斯坦纳菌的下降程度与PD改善呈正相关(r=0.50,P<0.001)。结论:牙周洁刮治术后福赛斯坦纳菌的下降程度可以预测牙周洁刮治术的治疗结果。TaqMan实时荧光定量PCR法能用于牙周治疗疗效的评价。     

牙周组织病

失活剂泄漏致牙周组织坏死19例分析；福赛斯坦纳菌与牙周洁刮治疗效的关系；一次与分次完成牙周基础治疗的微生物学比较研究；雌激素受体XbaⅠ和PvuⅡ基因多态性与慢性牙周炎的相关研究；慢性牙周炎龈下菌斑中福赛斯坦纳菌的检测.     

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。     

牙龈卟啉单胞菌PG0717基因与慢性牙周炎的相关性研究

目的检测慢性牙周炎患者和牙周健康者龈下菌斑中牙龈卟啉单胞菌（P.gingivalis）PG0717基因,探讨PG0717基因与牙周临床指数之间的关系。方法选取慢性牙周炎（CP）患者90例和牙周健康者90例,共采集龈下菌斑标本540个;记录临床牙周指数（牙周探诊深度、临床附着丧失和探诊出血）;设计特异性引物检测P.gingivalis阳性龈下菌斑标本的PG0717基因。结果在P.gingivalis阳性龈下菌斑中,CP组PG0717基因检出率显著高于对照组,分别为56.22%和41.27%（掊2=4.50,P＜0.05）;随着牙周探诊深度、临床附着丧失加重和探诊出血趋势的增加,CP组该基因检出率呈现增高趋势。结论PG0717基因与P.gingivalis的致病性有关。     

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。     

应用实时荧光定量PCR定量检测慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌

目的 检测慢性牙周交患者和牙周健康人龈下菌斑中牙龈卟啉单胞菌、总菌量和牙龈卟啉单胞菌所占比例,探讨牙龈卟啉单胞菌与牙周炎发生发展的关系.方法 采集经常规PCR方法检测牙龈卟啉单胞菌为阳性的76例慢性牙周炎患者和25例牙周健康者的龈下菌斑,应用TaqMan实时荧光定量PCR方法定量检测样本中牙龈卟啉单胞菌、总菌量;构建含有牙龈卟啉单胞菌和真细菌扩增片断的重组质粒,建立定量标准.结果 本研究设计的引物和探针具有良好的特异性及敏感性.病变位点龈下菌斑中牙龈卟啉单胞菌数量和总菌量均比健康位点高(P<0.001),两组位点牙龈卟啉单胞菌在菌斑中的比例没有差异(P>0.05);细菌数量与探诊深度间存在显著正相关关系(P<0.001),不同的探诊深度牙龈卟啉单胞菌所占比例无统计学差异(P>0.05).结论 龈下菌斑中牙龈卟啉单胞菌的数量水平及细菌总量与牙周状况、牙周炎发展有密切关系,实时荧光定量PCR对牙周病学研究具有广泛的应用前景.     

Occurrence of Porphyromonas gingivalis and Tannerella forsythensis in periodontally diseased and healthy subjects

BACKGROUND: The purpose of this study was to compare the prevalence and level of Porphyromonas gingivalis (P. gingivalis) and Tannerella forsythensis (T. forsythensis) in subgingival plaque samples from both healthy individuals and periodontal patients in different age groups. METHODS: A total of 498 subgingival plaque samples were studied. These samples were collected from 407 individuals diagnosed with periodontal disease (210 adult periodontitis [AP], 78 rapidly progressive periodontitis [RPP], and 119 refractory periodontitis [Ref-P] cases) and 91 healthy (H) subjects. P. gingivalis and T. forsythensis were detected by indirect immunofluorescent assay using species-specific polyclonal antisera to P. gingivalis strain (FDC 381) and T. forsythensis strain (FDC 335). The prevalence of P. gingivalis and T. forsythensis was compared by chi-square analysis. Differences in P. gingivalis and T. forsythensis levels among various periodontal status and age groups was determined by one-way analysis of variance and Fisher's multiple comparison tests. The association between the presence of P. gingivalis or T. forsythensis in different periodontal status and age groups was measured using odds ratios. RESULTS: P. gingivalis was found in 85.7% (P < 0.0001) and T. forsythensis in 60.7% (P = 0.0002) of diseased subjects compared to 23.1% and 39.6%, respectively, in healthy subjects. P. gingivalis, but not T. forsythensis, was detected more frequently in any diseased group than in the H group in every age group (P<0.0001). No significant difference was found in the prevalence of P. gingivalis and T. forsythensis among age groups, except T. forsythensis was more prevalent in the age group of 40 to 59 years than in the age group < 20 years (chi2 = 3.93, P = 0.047) in the H group. The mean level of P. gingivalis and T. forsythensis was significantly higher in diseased groups than in the H group (P < 0.0001). The odds ratio for P. gingivalis in the AP group (25.0) was greater than any other group for P. gingivalis or T. forsythensis compared to the H group. CONCLUSIONS: These data suggested that P. gingivalis is closely associated with the pathogenesis of periodontitis and that it may not be a normal inhabitant of periodontally healthy subjects. T. forsythensis is also important in the pathogenesis of periodontal disease; however, whether it causes periodontal disease or is a secondary invader of periodontal lesions remains to be determined.     

Detection of Tannerella forsythensis (Bacteroides forsythus) and porphyromonas gingivalis by polymerase chain reaction in subjects with different periodontal status

BACKGROUND: The aim of this study was to determine the prevalence of Tannerella forsythensis (formerly Bacteroides forsythus) and Porphyromonas gingivalis in subgingival plaque samples by using polymerase chain reaction (PCR), and to assess the relationship of these bacteria with different categories of periodontal disease and health. METHODS: Subjects were distributed into 3 groups according to their periodontal diagnosis: group 1, periodontally healthy (N = 10); group 2, periodontitis with probing depth < or = 5 mm (N = 10); group 3, periodontitis with probing depth > 5 mm (N = 10). The subjects in groups 2 and 3 had healthy and diseased periodontal sites. Subgingival plaque samples were obtained using paper points inserted into periodontal pockets (diseased sites) and into healthy gingival sulci (healthy sites) of the same subject. RESULTS: The distribution of bacteria differed in healthy and diseased sites. T. forsythensis (B. forsythus) was not detected in any sample from healthy sites in any group but was detected in 70% and 100% of diseased sites in groups 2 and 3, respectively. P. gingivalis was detected in only one sample from a healthy site (group 2), and in the diseased sites, its prevalence was 40% (group 2) and 90% (group 3). In addition, T. forsythensis (B. forsythus) and P. gingivalis were both detected in 30% and 90% of the diseased sites in groups 2 and 3, respectively. CONCLUSION: These results indicate a possible association between periodontal disease and the presence of T. forsythensis (B. forsythus) and/or P. gingivalis.     

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.     

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。     

Persistent colonization with Tannerella forsythensis and loss of attachment in adolescents

Colonization with Tannerella forsythensis may characterize the conversion of periodontally healthy sites into diseased sites. This three-year study describes the prevalence of T. forsythensis and its relationship to clinical loss of attachment (LOA) in a group of adolescents considered at risk of developing early chronic periodontitis. Adolescents with (LOA+) and without (LOA-) loss of attachment were examined at baseline and 1.5 and 3 yrs subsequently. On each occasion, attachment loss was measured on selected teeth, and the presence of T. forsythensis in their subgingival plaque samples was determined by PCR. T. forsythensis prevalence in LOA+ subjects at baseline (64%) increased to 82% and 86% on subsequent examinations. In contrast, prevalence of T. forsythensis in LOA- subjects was always significantly lower (25%, 36%, and 32%, respectively). The odds of loss of attachment were 8.16 times greater in subjects infected with T. forsythensis at each examination. These results suggest that T. forsythensis is strongly associated with loss of attachment in this adolescent population.     

Identification of periodontopathic bacteria in gingival tissue of Japanese periodontitis patients

INTRODUCTION: The identification of invading periodontopathic bacteria in tissues is important to determine their role in the pathogenesis of periodontal disease. The objective of this study was to identify periodontopathic bacteria in diseased gingival tissue of periodontitis patients. METHODS: Subgingival plaque and gingival tissue were collected from 32 generalized chronic periodontitis (CP), 16 generalized aggressive periodontitis (GAgP) and eight localized aggressive periodontitis (LAgP) patients. Detection frequencies and quantities of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Tannerella forsythensis were investigated by polymerase chain reaction. The prevalences of Streptococcus oralis and Streptococcus sobrinus were also examined and the distribution of A. actinomycetemcomitans serotypes was observed. RESULTS: P. gingivalis and T. forsythensis were detected in approximately 70% of tissue samples and 50% of plaque samples in the three periodontitis groups. Prevalence of A. actinomycetemcomitans in tissue samples was higher in the LAgP (63%) group than in either the CP (16%) or the GAgP (38%) group. A. actinomycetemcomitans serotype c was detected in 50% of LAgP patients. Detection frequencies of S. oralis and S. sobrinus were markedly low in both plaque and tissue samples from all three periodontitis groups. Amounts of P. gingivalis, A. actinomycetemcomitans and T. forsythensis in the tissue samples were not different among the three periodontitis groups. CONCLUSION: P. gingivalis, A. actinomycetemcomitans and T. forsythensis can localize in diseased gingival tissue and may be involved in periodontal tissue destruction. Serotype c is the predominant serotype of A. actinomycetemcomitans in Japanese LAgP patients.     

Clinical and microbiological profiles of human immunodeficiency virus (HIV)-seropositive Brazilians undergoing highly active antiretroviral therapy and HIV-seronegative Brazilians with chronic periodontitis

BACKGROUND: This study compares the periodontal clinical profile and the composition of the subgingival microbiota of human immunodeficiency virus (HIV)-seropositive and HIV-seronegative subjects with chronic periodontitis. METHODS: A total of 172 subjects were distributed into two HIV-seropositive groups (37 chronic periodontitis [H+CP+] and 35 periodontally healthy [H+CP-] individuals) and two HIV-seronegative groups (49 chronic periodontitis [H-CP+] and 51 periodontally healthy [H-CP-] subjects). Subgingival samples were collected from six sites with the deepest probing depth in the periodontitis groups and six random sites in the groups with periodontal health. All HIV-infected patients had undergone highly active antiretroviral therapy (HAART) for at least 2 years. The presence and levels of 33 bacterial species were detected by DNA probes and the checkerboard method. Kruskal-Wallis and Mann-Whitney tests were used to seek for significant differences among and between groups. RESULTS: H-CP+ patients showed significantly more periodontal destruction and inflammation than H+CP+ patients, whereas H+CP- subjects presented a greater percentage of sites with bleeding than H-CP- subjects (P <0.01). Patients who were HIV seronegative showed higher prevalence and levels of most bacterial species than HIV seropositive patients. Periodontal pathogens including Tannerella forsythensis, Porphyromonas gingivalis, Prevotella nigrescens, Eubacterium nodatum, Fusobacterium nucleatum, and Selenomonas noxia were more frequently detected in H-CP+ subjects compared to H+CP+ and controls. In contrast, Enterococcus faecalis and Acinetobacter baumannii were more commonly found in HIV-infected than in non-HIV-infected subjects (P <0.05). CONCLUSION: Putative periodontal pathogens are more prevalent in the subgingival microbiota of HIV-seronegative patients with chronic periodontitis, whereas species not usually associated with periodontitis are detected in higher frequency in HIV-seropositive subjects under HAART.     

Detection of herpetic viruses in gingival crevicular fluid of patients suffering from periodontal diseases: prevalence and effect of treatment

Background/Aim:  Although the role of bacteria in the etiology of periodontitis is well established, it has been suggested that herpetic viruses could contribute to the initiation and progression of this disease. The aim of this study was to determine the prevalence of human cytomegalovirus (HCMV), Epstein–Barr virus (EBV) and herpes simplex virus (HSV) in gingival crevicular fluid (GCF) samples obtained from periodontally healthy, gingivitis and periodontitis patients. In addition, the effect of periodontal treatment (scaling and root planing) on the persistence of herpetic viruses was evaluated in a sub-group of patients suffering from chronic periodontitis.
Methods:  The presence of viruses in GCF samples was assessed by a nested PCR amplification technique. The persistence of viruses in periodontal sites was evaluated following a scaling and root planing therapy.
Results:  A statistically significant higher prevalence of HCMV was observed in periodontitis patients as compared to healthy control subjects (35 vs. 8%, respectively; P  = 0.0377). A trend for a higher prevalence of HSV was also noted in the periodontitis group, in comparison with healthy control subjects. In addition, a higher prevalence of HCMV was associated with deep periodontal pockets in subjects suffering from periodontitis. In the sub-group of periodontitis patients, periodontal therapy resulted in the elimination (HCMV and EBV) or reduction (HSV) of the herpetic viruses.
Conclusions:  This study showed that the prevalence of HCMV and HSV viruses in GCF is higher in patients suffering from periodontitis compared to periodontally healthy subjects, and that the prevalence of HCMV is higher in deep periodontal pockets. It also brought evidences that periodontal therapy may be associated with virus elimination in diseased sites.