PCR直接检测龈下菌斑主要可疑牙周致病菌

目的：应用PCR方法直接检测龈下菌斑主要可疑牙周致病菌与牙周病活动部位的关系,探讨其方法的可行性并探讨其主要可疑牙周致病菌的分布规律。方法：应用聚合酶链反应(polymerase chain reaction,PCR)直接检测龈下菌斑主要可疑致病菌16s RNA保守区域片段。40名受试者包括牙周病患者20人,每人同口取一个牙周病活动部位,一个相对健康或牙周病静止对照部位;成人健康者20人,每人各取一个标本。结果：龈下菌斑5种可疑牙周致病菌在牙周病活动部位的检出率牙龈卟啉菌为86％,福赛类杆菌为95％,螺旋体为86％,中间普氏菌和黑色普氏菌分别为95％和33％,均显著高于同口部位对照组和健康对照组。结论：PCR直接检测菌斑牙龈卟啉单胞菌、中间普氏菌、福赛类杆菌、齿密螺旋体及黑色普氏菌匀与牙周炎活动部位相关。     

正常口腔中的可疑牙周致病菌

本实验对49例正常口腔的龈上和龈下菌斑内的可疑牙周致病菌(SPB)的分布情况进行了观察分析。 49例研究对象男21例,女28例;6～25岁,无龋,无牙周病的健康人。三月内未服用抗生素,未接受任何牙周治疗。定对在右上颌第一磨牙(No.3)的龈上及龈下菌斑。用BHI培养基厌氧培养37℃,4天以后进行细菌菌落计数,转种分离和鉴定。龈上菌斑内SPB的检出率按其高低顺序依次为:二氧化碳噬纤维菌、梭杆菌、消化链球菌、唾液弯曲杆菌、产黑色素类杆菌、优杆菌,溶蚀艾肯氏菌、生疾月形单胞菌。二氧化碳噬纤维菌中依次为牙龈二氧化碳噬纤维菌、黄褐二氧化碳噬纤维菌及生疾二氧化碳噬纤     

口源性口臭患者主要厌氧菌的分布

目的分析口臭患者龈下菌斑和舌苔上主要相关厌氧菌的分布情况。方法选择口腔门诊中口臭患者29例,鼻闻法来确定产臭部位和非产臭部位;分别采集龈下菌斑和舌苔标本接种在非选择性培养基和核梭杆菌选择性培养基,厌氧培养5d后记录非选择性培养基上生长的细菌总数、产黑色素细菌总数及核梭杆菌选择性培养基上生长的目的菌总数。结果29例患者中,15例患者的口臭主要来源于龈缘菌斑,10例主要来源于舌苔,4例患者的口臭由龈缘菌斑和舌苔共同产生;产臭部位和非产臭部位相比,细菌总数、产黑色素菌和具核梭杆菌数都明显上升（P〈0．01）。结论口源性口臭患者口气变化与产黑色素细菌、核梭杆菌相关。     

青少年牙周炎细菌病因学研究

应用选择和非选择性培养基的方法,采集了43名青少年牙周炎患者,31名龈炎患者和13名牙周健康者的303个龈下菌斑标本进行了放线共生嗜血杆菌、嗜二氧化碳噬纤维菌和产黑色素类杆菌的分离培养。结果表明:放线共生嗜血杆菌、嗜二氧化碳噬纤维菌在青少年牙周炎组的检出率显著高于其它     

二氧化碳噬纤维菌在牙周炎、牙龈炎患者及健康人口腔中分布的研究

为了解Capno菌与牙周病的关系,本实验分别对25例成人牙周炎、17例牙龈炎患者以及153名健康人(36名儿童,34名青年,49名老年人及34名与牙周病患者对照研究的健康成人)的龈上菌斑、龈下菌斑和唾液标本中的Capno菌的检出结果进行了统计学分析。研究表明:Capno菌在牙周炎、牙龈炎患者及健康人的菌斑及唾液中的检出率无差异(P>0.05),认为该菌是口腔的正常菌群之一。Capno在健康人菌斑中的检出率以儿童、青年为高,可能是青少年口腔中的优势菌群之一。     

牙周炎患者龈下厌氧菌群的分离和鉴定

从34例成年人牙周炎和18例青少年牙周炎患者及24例正常人的龈下菌斑中分离并鉴定了26种共311株革兰氏阴性无芽胞厌氧菌,其中产黑素类杆菌、牙龈类杆菌、伴放线嗜血杆菌、具核梭杆菌、衣氏放线菌和梅氏放线菌是牙周炎患者龈下优热菌群,小韦荣氏球菌的检出率与正常人比较无显著性差异。实验结果提示牙周炎为非单一的病原体所引     

替硝唑在牙周炎治疗中的临床及微生物学观察

本文通过对３０例牙周炎患者在口服替硝唑前后的临床症状及龈下菌斑的暗视野显微镜下螺旋体等菌种比例的变化,观察该药对牙周炎及其致病菌的疗效。结果：服用替硝唑后,在龈下菌斑中螺旋体的比例明显下降,而球菌的比例明显增高。牙龈指数、牙龈出血指数和探针深度均较服药前有明显好转（Ｐ＜０．００１）。提示：替硝唑能有效地抑制和杀灭龈下菌斑中的厌氧菌,并迅速消除牙周炎症,减轻症状。     

牙周炎患者菌斑细菌的药敏分析

目的:了解牙周炎患者龈下菌斑细菌对于口腔常用抗菌药物的敏感性.方法:从慢性牙周炎患者龈下菌斑中分离培养牙周可疑致病菌,测定6种口腔常用抗菌药物对其的最低抑菌浓度,分析以上致病菌对这几类抗菌药物的敏感性.结果:共从50名慢性牙周炎患者牙周袋中分离培养72株革兰阴性厌氧菌进行药物敏感试验.全部实验菌株都对阿莫西林/克拉维酸敏感.对甲硝唑、替硝唑、奥硝唑未见耐药株产生.结论:对于慢性牙周炎患者龈下菌斑中分离的厌氧菌,阿莫西林/克拉维酸和硝基咪唑类药物甲硝唑、替硝唑和奥硝唑的抑菌作用强.     

血链球菌在不同牙周状态下的分布及相关研究

目的：探讨口腔主要过氧化氢产生菌血链球菌和口腔链球菌在不同牙周健康状态下龈下菌群中的分布,及与牙周健康状态和牙龈卟啉单胞菌群中分布的相互关系。方法：纳入符合标准的受试者30人,受试位点86个,其中健康组11人,位点30个,龈炎组9人,位点29个,慢性牙周炎组10人,位点27个,检查记录牙周健康状态[包括牙龈指数（GI）和牙周袋深度（PD）],采集龈下菌斑标本,经厌氧菌培养基和AP-PCR及PCR鉴定后,将各受试组进行比较分析。结果：共获得草绿色链球菌523株,产黑色素菌241株。经AP-PCR及PCR鉴定后,得到血链球菌112株,口腔链球菌56株,牙龈卟啉单胞菌84株,健康组龈下菌斑中血链球菌,口腔链球菌和牙龈卟啉单胞菌构成比与牙周炎组相比有差异显著性;血链球菌和口腔链球菌与GI、PD呈负相关,牙龈卟啉菌与GI、PD呈正相关;血链球菌的构成与牙龈卟啉单胞菌的构成比呈负相关。结论：血链球菌等过氧化氢产生菌在龈下菌斑中比例的下降。可能是微生态失衡,致病菌过度增殖的重要机制。     

牙龈卟啉单胞菌、嵴链球菌、口腔链球菌属在口腔内不同解剖部位的分布

目的探讨牙龈卟啉单胞菌(Porphyromonas gingivalis)、嵴链球菌(Streptococcus cristatus)、口腔链球菌属(Streptococci oralis)在慢性牙周炎患者及牙周健康者不同口腔解剖部位生物膜的分布情况。方法选取慢性牙周炎患者25例,牙周健康者24例,分别作为慢性牙周炎组及健康对照组。测量临床指标(探诊深度、附着丧失和探诊出血),取受试者龈下菌斑、舌背、颊黏膜和唾液样品。Real-time PCR分析受试者不同受检部位S.cristatus、P.gingivalis、Streptococci oralis相对数量。结果慢性牙周炎组四个受检部位中P.gingivalis数量均大于健康对照组;慢性牙周炎组龈下菌斑中P.gingivalis数量大于其余受检部位;而慢性牙周炎组龈下菌斑、舌背、颊黏膜三个受检部位S.cristatus、Streptococci oralis数量小于健康对照者。结论与牙周健康者比较,慢性牙周炎患者口腔内不同解剖位置P.gingivalis数量增多,S.cristatus、Streptococci oralis数量减少;P.gingivalis检出数量增加提示牙周炎患病风险增加,而S.cristatus、Streptococci oralis检出数量降低提示牙周炎患病风险降低。     

Growth, incidence and activities of dissimilatory sulfate-reducing bacteria in the human oral cavity

Abstract Viable counts and activities of sulfate-reducing bacteria were determined in the oral cavities of 12 healthy volunteers. Of these, 10 harboured viable sulfate-reducing bacteria populations. Six separate sites were sampled: the posterior tongue, anterior tongue, mid buccal mucosa, vestibular mucosa, supragingival plaque and subgingival plaque. Sulfate-reducing bacteria occurred in all areas, with the highest incidence in supragingival plaque. Viable counts and sulfate-reducing activities in each of the regions varied from 0 to 10⁸ cfu (g wet weight)⁻¹ and from 0 to 50 nmol (g wet weight) ⁻¹ h⁻¹, respectively. As sulfate-reducing bacteria can be detected in the oral cavity, they may potentially be involved in terminal oxidative processes carried out by the microflora of the mouth.     

Growth and Activities of Sulfate-Reducing and Methanogenic Bacteria in Human Oral Cavity

Viable counts and activities of sulfate-reducing bacteria (SRB) and methanogenic bacteria were determined in the oral cavities of eight volunteers. Of these, seven harbored viable SRB populations, and six harbored viable methanogenic bacterial populations. Two volunteers classified as type III periodontal patients had both SRB and methanogenic bacteria. Six separate sites were sampled: posterior tongue, anterior tongue, mid-buccal mucosa, vestibular mucosa, supragingival plaque, and subgingival plaque. The SRB was found in all areas in one volunteer, and it was mostly present in posterior tongue, anterior tongue, supragingival, and subgingival plaques in many volunteers. The methanogenic bacteria were mostly found in supragingival and subgingival plaques. The activities of sulfate reduction and methane production were determined in randomly selected isolates. Received: 27 July 2002 / Accepted: 27 August 2002     

Relationship of periodontal clinical parameters with bacterial composition in human dental plaque

More than 600 bacterial species have been identified in the oral cavity, but only a limited number of species show a strong association with periodontitis. The purpose of the present study was to provide a comprehensive outline of the microbiota in dental plaque related to periodontal status. Dental plaque from 90 subjects was sampled, and the subjects were clustered based on bacterial composition using the terminal restriction fragment length polymorphism of 16S rRNA genes. Here, we evaluated (1) periodontal clinical parameters between clusters; (2) the correlation of subgingival bacterial composition with supragingival bacterial composition; and (3) the association between bacterial interspecies in dental plaque using a graphical Gaussian model. Cluster 1 (C1) having high prevalence of pathogenic bacteria in subgingival plaque showed increasing values of the parameters. The values of the parameters in Cluster 2a (C2a) having high prevalence of non-pathogenic bacteria were markedly lower than those in C1. A cluster having low prevalence of non-pathogenic bacteria in supragingival plaque showed increasing values of the parameters. The bacterial patterns between subgingival plaque and supragingival plaque were significantly correlated. Chief pathogens, such as Porphyromonas gingivalis, formed a network with other pathogenic species in C1, whereas a network of non-pathogenic species, such as Rothia sp. and Lautropia sp., tended to compete with a network of pathogenic species in C2a. Periodontal status relates to non-pathogenic species as well as to pathogenic species, suggesting that the bacterial interspecies connection affects dental plaque virulence.     

Sanger和Pyrosequencing测序法分析健康成人口腔菌群

目的对比Sanger和Pyrosequencing测序法分析健康人口腔菌群组成。方法收集6例健康成人唾液、舌背、黏膜、龈上及龈下菌斑并构建16SrRNA基因文库,分别用Sanger和Pyrosequencing测序法分析。结果 Sanger测序所得已知的序列有5,794条(占6,535总序列数88.7%)、75个属,396个序列划分操作分类单元(operational taxonomic units,OTUs,占总OTUs的61.4%)。Pyrosequencing测序所得已知的序列有10,771条(占11,103总序列数97.0%)、66个属,322个OTUs(占总OTUs的68.0%)。Sanger和Pyrosequencing测序法所得口腔菌群在门、属的水平分布趋势基本一致,但在种的水平分布差异显著。Sanger和Pyrosequencing测序法构建的口腔菌群文库均匀度值分别为0.016和0.007,说明Pyrosequencing分析口腔菌群物种数量分布比Sanger测序方法的文库均匀性稍差,但优势种更显著。结论 Pyrosequencing测序时所构建基因文库能代表口腔菌群的多样性且经济、省时,可以应用于口腔细菌物种的分析。     

牙周炎治疗过程中龈下菌斑的动态观察

目的　通过对牙周炎治疗过程中龈下菌斑内螺旋体和球菌的百分比组成的改变进行动态观察研究,为牙周炎的诊断和疗效评价提供依据。方法　选择１８ 例经门诊确诊为成人牙周炎的患者,随机分为Ａ、Ｂ、Ｃ 三组,均施以龈上洁治、龈下刮治和根面平整术。其中Ａ 组和Ｂ 组分别给予艾利克及洗必太含漱液含漱。所有患者每周复诊一次,采集龈下菌斑,进行刚果红染色,镜检计数螺旋体及球菌的百分比,同时检测患处牙龈指数（ＧＩ） 。结果　Ａ、Ｂ 组患者经治疗１ 周后,ＧＩ值变化极为显著（Ｐ＜ ０．０１） ,螺旋体的减少及球菌的增加也极具显著性（Ｐ＜ ０．０１） ,至３ 周时趋于稳定;Ｃ 组则在１ 周时ＧＩ值及菌斑组成的变化有显著性（Ｐ＜ ０．０５） ,至２ 周时变化才最为明显（Ｐ＜ ０．０１） ,４ 周时趋于稳定。结论　牙周基础治疗能明显改善临床症状,使螺旋体百分比显著降低,球菌百分比显著增加。口腔含漱液作为一种牙周炎的辅助用药,可明显改善菌斑的组成,促进正常口腔卫生环境的建立。     

Coaggregation properties of human oral Veillonella spp.: relationship to colonization site and oral ecology.

The primary habitats of oral veillonellae are the tongue, dental plaque, and the buccal mucosa. Isolates were obtained from each habitat and tested for coaggregation with a battery of other oral bacterial strains. All 59 tongue isolates tested for coaggregation were Veillonella atypica or Veillonella dispar. All but one of them coaggregated with strains of Streptococcus salivarius, a predominant inhabitant of the tongue surface but not subgingival dental plaque. These tongue isolates were unable to coaggregate with most normal members of the subgingival flora such as Actinomyces viscosus, Actinomyces naeslundii, Actinomyces israelii, and Streptococcus sanguis. In contrast, 24 of 29 Veillonella isolates, of which 20 were Veillonella parvula from subgingival dental plaque samples, coaggregated strongly with the three species of Actinomyces, S. sanguis, and other bacteria usually present in subgingival plaque, but they did not coaggregate with S. salivarius. The majority of isolates from the buccal mucosa (42 of 55) has coaggregation properties like those from the tongue. These results indicate that the three human oral Veillonella species are distributed on oral surfaces that are also occupied by their coaggregation partners and thus provide strong evidence that coaggregation plays a critical role in the bacterial ecology of the oral cavity.     

Coaggregation properties of human oral Veillonella spp.: relationship to colonization site and oral ecology

The primary habitats of oral veillonellae are the tongue, dental plaque, and the buccal mucosa. Isolates were obtained from each habitat and tested for coaggregation with a battery of other oral bacterial strains. All 59 tongue isolates tested for coaggregation were Veillonella atypica or Veillonella dispar. All but one of them coaggregated with strains of Streptococcus salivarius, a predominant inhabitant of the tongue surface but not subgingival dental plaque. These tongue isolates were unable to coaggregate with most normal members of the subgingival flora such as Actinomyces viscosus, Actinomyces naeslundii, Actinomyces israelii, and Streptococcus sanguis. In contrast, 24 of 29 Veillonella isolates, of which 20 were Veillonella parvula from subgingival dental plaque samples, coaggregated strongly with the three species of Actinomyces, S. sanguis, and other bacteria usually present in subgingival plaque, but they did not coaggregate with S. salivarius. The majority of isolates from the buccal mucosa (42 of 55) has coaggregation properties like those from the tongue. These results indicate that the three human oral Veillonella species are distributed on oral surfaces that are also occupied by their coaggregation partners and thus provide strong evidence that coaggregation plays a critical role in the bacterial ecology of the oral cavity.     

口腔卫生预防措施对固定正畸患者牙周菌群和牙周临床指数的影响

目的 采用牙周洁治和强化的口腔健康教育方法,通过检测牙周菌群和牙周临床指数的动态变化,探讨口腔卫生预防措施对固定正畸患者牙周菌群的影响。方法 选择正畸初诊患者20例,平均分为试验组（T组）和对照组（C组）。于正畸治疗开始前检查右上颌第一磨牙16和右下颌中切牙41牙周状况,并采集其近中颊轴嵴处龈缘菌斑做细菌分离培养。T组于治疗前进行全口洁治,每月复诊加力时均给予口腔卫生检查,强调口腔卫生的重要性;C组仅在初诊时进行口腔卫生指导,其余不作处理。2组患者分别于矫治器安装后1、3和6个月进行临床及细菌学检查。结果 随观察时间的延长,T组颊侧菌斑指数和牙龈指数在第1、3、6个月时较基线降低;C组颊侧菌斑指数第6个月时较基线降低,舌侧探诊深度则升高（P＜0.05）。细菌检出率和检出量的变化在T组可见韦荣球菌属降低而弯曲杆菌属和Gn产黑色素厌氧杆菌（BPAR）升高,C组消化链球菌属和BPAR升高（P＜0.05）;BPAR在第3个月、消化链球菌属在第6个月时T组检出率低于C组（P＜0.05）,而细菌检出量和牙周临床指数在2组间没有观测到处理因素的作用（P＞0.05）。结论 正畸前即存在牙龈炎的患者,建议进行预防性牙周洁治;牙周洁治必须和口腔卫生教育、正确的日常菌斑控制措施结合进行。     

应用PCR与温度梯度凝胶电泳分析龈上菌斑微生物群落

目的应用PCR与温度梯度凝胶电泳(PCR—TGGE)分子技术对成年健康口腔的龈上菌斑中微生物群落组成进行分析。方法8例成年个体包括4男4女,年龄19～29岁,分别采取每例个体上下颌牙周龈上菌斑样品,共18份(个体Subl间隔10天采集2次样品)。提取菌斑DNA,PCR扩增16SrDNAV3可变区,产物经TGGE后进行相似系数分析。结果同一个体的上下颌微生物群落组成相似性系数为81％～95％,而不同个体的龈上菌斑微生物群落组成相似性系数,均在60％以下。结论不同个体具有其独特的牙周微生物群落,而且在一定时期内组成稳定。