牙龈卟啉单胞菌毒力岛基因检出与慢性牙周炎的相关性研究

目的:检测慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌(Porphyromonas gingivalis)毒力岛中PG0836、PG0838和PG0839基因,探讨3个基因与临床牙周指数之间的关系.方法:选取慢性牙周炎(chronic periodoiltitis,CP)患者90例,共采集龈下菌斑标本270个.记录受试位点的牙周探诊深度、临床附着丧失和探诊出血情况.设计特异性引物,检测P.gingivalis阳性龈下菌斑标本的PG0836、PG0838和PG0839基因.采用SPSS 11.0软件包进行统计学分析,采用X2检验对不同牙周临床指数基因的检出率进行比较.结果:在CP患者P.gingivalis阳性龈下菌斑中,PG0836、PG0838和PG0839基因检出率分别为66.17%、24.88%和27.86%.探诊出血阳性位点的PG0839基因检出率(28.35%)显著高于探诊出血阴性位N(14.29%,P<0.05).在牙周探诊深度为4～6mm和>6mm的受试位点,PG0839基因检出率均显著高于牙周探诊深度<4mm的位点(P<0.05).PG0836和PG0838基因在不同牙周指数组检出率无显著差异.结论:PG0839基因的检出与CP病损部位的临床指标呈正相关关系,提示该基因可能与P.gingivalis的致病性有关.     

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。     

伴放线菌嗜血菌在慢性牙周炎患者和牙周健康者中的分布

目的分析伴放线菌嗜血菌在慢性牙周炎患者和牙周健康者中的分布情况。方法选择116例慢性牙周炎患者（CP组）和111例牙周健康者（健康组）为研究对象。CP组选取磨牙区牙周袋最深的2个患牙的探诊深度最深点作为患牙的取样位点,同时选取磨牙或前磨牙区1个健康牙的近中位点作为健康牙的取样位点;健康组选取上颌第一磨牙近中位点作为取样位点。收集取样位点的龈下菌斑,提取DNA,用16S rRNA聚合酶链反应检测伴放线菌嗜血菌的分布;同时检查并记录取样牙的牙周临床指数（包括探诊深度、临床附着丧失和探诊出血）,分析伴放线菌嗜血菌检出率和牙周临床指数的关系。结果CP组患牙伴放线菌嗜血菌检出率为33.62%,明显高于CP组健康牙和健康组（P<0.01）。伴放线菌嗜血菌检出率随着患者年龄的增加而降低,随着探诊深度、临床附着丧失的增加而增加,探诊出血阳性患牙的检出率（37.07%）也明显高于探诊出血阴性的患牙（7.41%）（P<0.05）。结论伴放线菌嗜血菌检出率随着牙周炎症程度的加重而升高,与慢性牙周炎的发生发展具有密切的联系。     

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。     

不同牙周状况龈下菌斑中福赛斯坦纳菌分布

目的：分析牙周健康者和慢性牙周炎病人龈下菌斑中福赛斯坦纳菌的分布情况,探讨该菌与不同牙周状况的关系。方法：收集111例牙周健康者、108例慢性牙周炎病人,病变位点和健康位点共327个龈下菌斑,采用16S rRNA PCR方法检测福赛斯坦纳菌的分布。结果：牙周炎病人病变位点福赛斯坦纳菌检出率为56.5%,显著高于牙周健康者（10.8%,P〈0.001）。慢性牙周炎病人健康位点福赛斯坦纳菌检出率为3.7%,显著低于病变位点（P〈0.001）,牙周健康者该菌的检出率比牙周炎健康位点高（P〈0.05）。结论：龈下菌斑中福赛斯坦纳菌的存在与牙周炎症的发生有密切关系。     

Ⅱ型糖尿病合并牙周炎患者龈下菌斑微生物群落分析

目的：本研究旨在比较Ⅱ型糖尿病合并慢性牙周炎患者与无糖尿病的牙周炎患者龈下菌斑微生物构成。方法：12名患者分为糖尿病合并慢性牙周炎组（T2DM＋CP组）与慢性牙周炎组（CP组）2组,各6人。记录所有患者基本信息及牙周临床参数,包括年龄、性别、探诊深度和附着丧失。根据探诊深度和附着丧失取患病位点的菌斑样本。PCR检测7种牙周可疑致病菌。采用DGGE分离扩增的16SrDNA片段。结果：两组结果显示两组牙周参数无显著差异。两组7种细菌检出率相似。所有对象中均检出牙龈卟啉单胞菌、福塞坦氏菌、齿垢密螺旋体和中间普氏菌,而具核梭杆菌在两组中均有一个样本未检出。变黑普氏菌在T2DM＋CP组的2个样本中检出,而CP组有4个样本检出。伴放线共聚菌在所有样本中均未检测到。DGGE分析结果示两组间条带数量及树状聚类分析均无显著差异。结论：Ⅱ型糖尿病合并牙周炎患者龈下菌斑的牙周可疑致病菌的检出情况以及DGGE分析与无糖尿病患者相似。     

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。     

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。     

牙周袋内挥发性硫化物与牙周炎症程度的关系

目的初步探讨牙周袋内挥发性硫化物与侵袭性牙周炎(aggressive periodontitis,AgP)和慢性牙周炎(chronic periodontitis,CP)炎症程度的关系。方法用金刚牙周探针诊断仪检查探诊深度、附着丧失、探诊出血等临床指标的同时,检测牙周袋内硫化物水平。共检查15例AgP和16例CP患者870个患牙的5 220个位点。结果无论是AgP还是CP患者,硫化物阳性位点的各项临床指标均明显高于阴性位点(P<0.001);硫化物水平与各项临床指标间都有明显的正相关关系(P<0.001);中、深袋组的硫化物水平和阳性位点率均明显高于浅袋位点(P<0.001);有附着丧失位点的硫化物水平和阳性位点率均高于无附着丧失位点,在浅袋组其差异有显著性。结论牙周袋内挥发性硫化物的检测可以反映侵袭性牙周炎和慢性牙周炎患者的牙周炎症程度。     

牙龈卟啉单胞菌与牙周病相关性的聚合酶链反应研究

目的利用PCR检测慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌(Porphyromonas gingivalis,P. g)的16S rDNA水平,通过检测该基因水平来探讨牙龈卟啉单胞菌的水平与牙周病的相关性.方法采集慢性牙周炎患者12例共36个龈下菌斑标本,记录临床指标(探诊深度、临床附着水平、牙龈指数、菌斑指数、龈沟出血指数),PCR检测龈下菌斑标本中的P. g 16S rDNA基因,扩增产物经琼脂糖电泳、拍照后,应用计算机软件GeneTools扫描并计量其中的相对核酸含量.结果P. g16S rDNA水平与探诊深度(P＜0.001)及牙龈指数(P＜0.01)之间存在正相关关系.结论P.g16S rDNA水平与牙周状态密切相关,对于P.g16S rDNA水平的监测有望成为牙周病诊断和治疗方案制定的辅助检查手段.     

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.     

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.     

Clinical and microbiological effect of scaling and root planing in smoker and non-smoker chronic and aggressive periodontitis patients

OBJECTIVES: To compare the effects of scaling and root planing (SRP) on clinical and microbiological parameters at selected sites in smoker and non-smoker chronic and generalized aggressive periodontitis patients. MATERIALS AND METHODS: Clinical parameters including probing depth (PD), relative attachment level (RAL), and bleeding upon probing (BOP), and subgingival plaque samples were taken from four sites in 28 chronic periodontitis (CP) and 17 generalized aggressive periodontitis (GAgP) patients before and after SRP. Polymerase chain reaction assays were used to determine the presence of A. actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia and Treponema denticola. RESULTS: Both CP and GAgP non-smokers had significantly greater reduction in pocket depth (1.0+/-1.3 mm in CP smokers versus 1.7+/-1.4 mm in non-smokers, p=0.007 and 1.3+/-1.0 in GAgP smokers versus 2.4+/-1.2 mm in GAgP non-smokers, p<0.001) than respective non-smokers, with a significant decrease in Tannerella forsythensis in CP sites (smokers 25% increase and non-smokers 36.3% decrease, p<0.001) and Prevotella intermedia at GAgP sites (smokers 25% reduction versus 46.9% in non-smokers, p=0.028). CONCLUSION: SRP was effective in reducing clinical parameters in both groups. The inferior improvement in PD following therapy for smokers may reflect the systemic effects of smoking on the host response and the healing process. The lesser reduction in microflora and greater post-therapy prevalence of organisms may reflect the deeper pockets seen in smokers and poorer clearance of the organisms. These detrimental consequences for smokers appear consistent in both aggressive and CP.     

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.     

Possible periodontal pathogens associated with clinical symptoms of periodontal disease in Japanese high school students

BACKGROUND: The purpose of this study is to investigate how the components of biofilm and clinical oral status change in adolescents and to identify specific periodontal pathogens as risk markers for the onset of periodontitis. METHODS: One hundred seven high school students (72 boys and 35 girls, all 15 years old) were recruited. The mesio-lingual site of the left lower first molar was selected as the examined site. Probing depth (PD), bleeding on probing (BOP), the presence of subgingival calculus, and Community Periodontal Index (CPI) were determined by examination with a WHO probe. The prevalence and proportion of seven selected periodontal pathogens (Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia, Campylobacter rectus, Eikenella corrodens, and Actinobacillus actinomycetemcomitans serotypes b and c) were determined by indirect immunofluorescent technique, and the prevalence and proportion of spirochetes were determined by their morphology under dark-field microscopy. The relationship between the periodontal status and the bacterial condition was statistically analyzed. RESULTS: The mean proportion of T. forsythensis was significantly higher in BOP (+) sites compared with BOP (-) sites (3.47% +/- 5.35% versus 0.83% +/- 1.95%) and in CPI 3 sites compared with CPI 0 sites (3.29% +/- 5.28% versus 0.68% +/- 1.37%). The mean proportion of C. rectus was significantly increased in BOP (+) compared with BOP (-) (2.01% +/- 2.48% versus 0.79% +/- 0.91%) and in CPI 3 sites compared with CPI 0 sites (2.04% +/- 2.64% versus 0.80% +/- 0.79%). CONCLUSION: The results indicated that T. forsythensis and C. rectus might be able to be used as risk markers for the onset of periodontitis.