武汉市306名儿童牙菌斑中变形链球菌的血清型

本试验共检测了306例5～6岁学龄前儿童牙菌斑中的变形链球菌,在其中208名患龋齿病儿童的牙菌斑中分离出304株变形链球菌,用双向琼脂扩散法进行血清学鉴定,c、e、f血清型有259株,占所分离的变链菌总数的85.19%,其中c型206株占68%,e型37株占12%,f型16株占5%。在有龋组和无龋组牙菌斑中变链菌培养阳性率比较,有龋组变链菌培养阳性率明显高于无龋组(P<0.01)。本试验提示变链菌是武汉地区龋齿病的重要致病因素之一;武汉地区龋齿病的儿童中感染的变形链球菌血清型以c型为主。     

感染根管的厌氧菌培养结果分析

从64只感染根管中的58只根管分离到144株无芽胞厌氧菌,其中类杆菌54株,厌氧性链球菌23株,韦荣氏球菌17株,真杆菌11株,梭杆菌10株,放线菌8株,双岐杆菌2株,消化链球菌和消化球菌19株。40只根管为厌氧菌和兼性厌氧菌或需氧菌混合感染,18只根管和6只根管分别为单独厌氧菌和兼性厌氧菌感染。33只根尖周炎根管分别采集牙髓和根尖渗出物样本进行培养,实验结果表明牙髓样本中革兰氏阳性厌氧杆菌检出率较高,根尖渗出物中以产黑素类杆菌属的细菌检出率较高。根尖周炎和牙槽脓肿患者的感染根管中产黑素类杆菌属的细菌检出率明显高于蜂窝组织炎患者。     

风化程度不同的黄金矿石中微生物的调查

对我国广西某地尚未开采的黄金矿床中风化程度不同的黄金矿石进行微生物分离,结果分离出6株霉菌和6株细菌,未发现酵母菌、放线菌和专性厌氧菌。三种风化程度不同的矿石中均分布有蜡状芽抱杆菌和氧化亚铁硫杆菌。     

小鼠盲肠内容物中正常菌群的分离和定量

本文报道了采用厌氧培养技术,从小鼠盲肠内容物中分离到6株严格厌氧和兼性厌氧菌。鉴定为拟杆菌、乳酸杆菌、双歧杆菌、难辨梭菌、粪链球菌和大肠扦菌。它们在小鼠盲肠内容物的正常菌群中占优势,含菌量与国外文献中报告的相似。     

健康青少年口腔正常菌群的初步研究

本文采用非选择性培养基对22名健康青少年的唾液、沟裂菌斑、龈上菌斑及龈下菌斑中的需氧菌、兼性厌氧菌及专性厌氧菌进行了分离培养,并计算其在不同标本中占可培养菌的百分比及检出率。结果共分离到包括18个菌属的35种细菌。其中,链球菌、放线菌、奈瑟氏球菌、二氧化碳噬纤维菌、类杆菌、梭杆菌,奴卡氏菌及棒状杆菌在口腔4个部位的检出率及所占比例均较高,是健康青少年口腔中的优势菌群.通过比较还发现,其中一些菌在口腔4个部位的分布存在一定差异.本文还采用刚果红负性染色涂片法,镜下观察龈上、龈下菌斑中的螺旋体,并计算其相对比例.结果龈下菌斑中螺旋体的相对比例明显高于龈上菌斑.     

汉族与维吾尔族儿童封闭型不同龋敏感中变形链球菌乳酸脱氢酶遗传多态性研究

目的探讨汉族与维吾尔族儿童封闭型不同龋敏感中变形链球菌乳酸脱氢酶活性与ldh基因多态性及龋病发生的关系。方法从前期实验分离的维吾尔族与汉族儿童口腔变形链球菌中筛选出:维吾尔族儿童封闭型口腔变形链球菌高龋组变形链球菌35株[龋失补牙数(dmft)≥5],无龋组变形链球菌37株[龋失补牙数(dmft)=0];27株表现乳酸脱氢酶高活性,24株低活性;汉族儿童口腔中分离的高龋和无龋变形链球菌各35株,高活性和低活性各25株,使用特异引物扩增细菌组DNA,获得结构基因ldh,进行聚合酶链反应-限制性内切酶多态性分析。结果维吾尔族儿童封闭型和汉族儿童MseⅠ酶切ldh-RFLP表现A、B两种基因型;而MnlⅠ、DdeⅠ、NlaⅢ和AluⅠ消化扩增产物未发现多态性。MseⅠ产生的B基因型菌株在维吾尔族儿童封闭型,汉族儿童高龋组的检出均高于无龋组(P0.05),A、B两种基因型在不同酶活性组的分布不同(P0.05),高酶活性组B基因型菌株的比例高于低酶活性组;A、B基因型在维吾尔族儿童封闭型和汉族高龋组、无龋组,高酶活性组和低酶活性组的检出比较差异无统计学意义(P0.05)。结论变形链球菌乳酸脱氢酶基因多态性在维吾尔族与汉族儿童并未表现出差异性,ldh基因多态性与变形链球菌乳酸脱氢酶活性及龋高敏感性相关。ldh基因分布与酶活性及乳牙龋具有正相关性。     

高龋菌株产生的变链素对口腔链球菌抑制活性的比较

目的 通过比较高龋和无龋儿童变形链球菌临床分离株产生的变链素对口腔链球菌的抑制活性,探讨变链素的产生与其致龋性及口腔微生态的关系.方法 从10例高龋儿童和10例无龋儿童牙菌斑内分离、鉴定得到80株变形链球菌临床分离株,分为高龋组和无龋组.用平板法检测两组菌株产生的变链素对口腔链球菌Streptococcus oralis ATCC 10557的抑制情况,观察两组菌株产生的抑菌环大小, 测量记录数据,T检验比较两组菌株抑菌环均数差异.结果 高龋组产生的抑菌环平均值为10.4 mm,无龋组平均值为6.9 mm,T检验显示差异具有显著性(t值为3.098,P<0.05).结论 高龋菌株产生的变链素对ATCC 10557有更大的抑制活性,变链素对口腔链球菌的抑制活性与其致龋力呈正相关.     

维吾尔族不同龋敏感儿童变形链球菌临床分离株表面蛋白V区遗传多态性研究

目的比较维吾尔族高龋和无龋儿童变链菌临床分离株表面蛋白V区遗传多态性与其合成水不溶性葡聚糖的关系。方法选取课题组前期实验所得的维吾尔族高龋儿童合成水不溶性葡聚糖能力较强的变形链球菌临床株18株和无龋儿童合成水不溶性葡聚糖能力较弱临床株12株。提取全菌DNA,经PCR扩增其表面蛋白可变区V区编码基因SrV~+后,利用限制性内切酶DdeⅠ进行限制性片段长度多态性分析。结果经DdeⅠ酶切后,高产糖组变链菌出现了4种基因型,低产糖组出现了3种基因型。这几种基因型在不同产糖组中的分布不同(P0.05)。结论维吾尔族不同龋敏感儿童变形链球菌临床分离株SrV~+基因的遗传多态性可能是其合成水不溶性葡聚糖能力出现差异的因素之一。     

天然药物蜂房化学成分提取物对口腔细菌生长的实验研究

目的研究蜂房中分离得到的不同组分对致龋菌生长的影响,寻找蜂房抑龋的有效成分。方法通过溶剂分段和层析技术对蜂房进行分离,得到4个组分,采用液体稀释法研究蜂房不同组分对口腔常居细菌——血液链球菌、唾液链球菌,以及4种主要致龋菌——变形链球菌、内氏放线菌、粘性放线菌和乳酸杆菌生长的影响,使用活菌计数法测量五倍子总鞣质及各组分对变形链球菌生长曲线的影响。结果蜂房提取物中1、2和3组分对实验菌有较强的抑菌作用,蜂房提取物3组分对于变形链球菌生长曲线的抑制作用最强。结论蜂房各组分对实验菌都有一定的抑菌作用,其抑菌作用可能和其中的甾醇类化合物有关。     

放线菌的基因型及其与根面龋发生的关系

根面龋的细菌学研究始于 2 0世纪 70年代 ,早期研究集中于革兰阳性丝状杆菌 ,尤其是黏性放线菌 (以下简称“黏放”)和内氏放线菌 (以下简称“内放”)。 80年代中后期 ,变链球菌在根面龋中的作用逐渐受到重视。目前认为根面龋是以变链球菌 ,放线菌和乳杆菌为主的混合感染。根面龋细菌学研究表明根面龋的菌群很复杂 ,在不同的病损阶段优势菌的种类呈现多样化[1 ,2 ] 。有学者通过对不同病变发展阶段的根面龋有关的微生物学研究 ,指出早期根面龋中主要是乳杆菌 ,内放菌和链球菌 ;中度根面龋中菌群较复杂 ,其中内放菌比链球菌更为常见 ,但未检出…     

Quantitative detection of Streptococcus mutans in the dental plaque of Japanese preschool children by real-time PCR

AIMS: To detect quantitatively the total bacteria and Streptococcus mutans in dental plaque by real-time PCR with prbac, Sm and GTF-B primers, and to compare their presence with the prevalence of dental caries in Japanese preschool children. METHODS AND RESULTS: Human dental plaque samples were collected from the labial surfaces of the upper primary central incisors of 107 children. The dental status was recorded as dft by WHO caries diagnostic criteria. Positive dt and dft scores by the Sm or GTF-B primer were significantly higher than negative scores (P < 0.01). The proportions of Strep. mutans to the total bacteria from sound, and sound and/or filled upper primary incisors were significantly lower than those from decayed or filled, and decayed incisors, respectively (P < 0.01). CONCLUSIONS: The ratios of Strep. mutans to total bacteria in plaque detected by real-time PCR with Sm and GTF-B primers were closely associated with the prevalence of dental caries in Japanese preschool children. SIGNIFICANCE AND IMPACT OF THE STUDY: These assays may be useful for the assessment of an individual's risk of dental caries.     

Streptococcus mutans and oral streptococci in dental plaque

The human oral microbial biota represents a highly diverse biofilm. Twenty-five species of oral streptococci inhabit the human oral cavity and represent about 20 % of the total oral bacteria. Taxonomy of these bacteria is complex and remains provisional. Oral streptococci encompass friends and foes bacteria. Each species has developed specific properties for colonizing the different oral sites subjected to constantly changing conditions, for competing against competitors, and for resisting external agressions (host immune system, physico-chemical shocks, and mechanical frictions). Imbalance in the indigenous microbial biota generates oral diseases, and under proper conditions, commensal streptococci can switch to opportunistic pathogens that initiate disease in and damage to the host. The group of "mutans streptococci" was described as the most important bacteria related to the formation of dental caries. Streptococcus mutans, although naturally present among the human oral microbiota, is the microbial species most strongly associated with carious lesions. This minireview describes the oral streptococci ecology and their biofilm life style by focusing on the mutans group, mainly S. mutans. Virulence traits, interactions in the biofilm, and influence of S. mutans in dental caries etiology are discussed.     

复方茶多酚含漱液对正畸儿童牙面菌斑中致龋菌的影响

目的观察复方茶多酚含漱液对正畸儿童牙面菌斑中细菌总数和变形链球菌数的影响,以及牙菌斑内原位pH的改变。方法选择42例戴用固定矫治器的正畸儿童,随机分为2组,试验组用复方茶多酚含漱液漱口,对照组用蒸馏水漱口。分别于戴用矫治器前,戴入后1月采集上下颌牙唇颊面菌斑,测定菌斑中细菌总数及变形链球菌数,同时测定牙菌斑原位pH。结果对照组戴用后1月,细菌总数及变形链球菌数较戴用前明显增加(P0.01),牙菌斑原位pH较戴用前降低(P0.01)。试验组与对照组戴用后1个月相比,试验组细菌总数及变形链球菌数明显少于对照组(P0.01),牙菌斑原位pH高于对照组(P0.01)。结论戴用固定矫治器后,牙面菌斑内细菌总数及变形链球菌数较戴用前增加,牙菌斑原位pH较戴用前降低,应用茶多酚含漱液可明显抑制正畸儿童口腔内变形链球菌数,减少龋坏发生。     

Evaluation of (GTG)<Subscript>5</Subscript>-PCR for rapid identification of <Emphasis Type="Italic">Streptococcus mutans</Emphasis>

Repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using the (GTG)(5) primer was applied for fast screening of bacterial strains isolated from dental plaque of early childhood caries (ECC)-affected children. A group of 29 Gram-positive bacteria was separated into a homogeneous cluster together with Streptococcus mutans reference strains and constituted an aberrant branch after the numerical analysis of (GTG)(5)-PCR fingerprints. Automated ribotyping with EcoRI restriction enzyme (RiboPrinter microbial characterization system) revealed high genetic heterogeneity among the tested group and proved to be a good tool for strain-typing purposes. Further characterization of the studied strains was achieved by extensive phenotyping and whole-cell protein fingerprinting and confirmed all the strains as S. mutans representatives. Obtained results showed rep-PCR fingerprinting with the (GTG)(5) primer to be a fast and reliable method for identification of S. mutans.     

甲壳胺影响变形链球菌粘附力的研究

目的：研究甲壳胺对变形链球菌的附着力的影响。方法：通过变链菌粘附实验和细菌平皿计数法，观察变链菌在有或无甲壳胺的培养基中的粘附和生长情况，结果：1．甲壳胺对变链菌的附着有抑制作用。2．甲壳胺能使已附着的变链菌脱落。0．5％浓度的甲壳胺作用强于0．25％。3．甲壳胺对变链菌的生长数量及形态无影响。结论：甲壳胺的防龋性是由于它减弱了变链菌的粘附力，而不是通过减少变链菌的数量来实现的。     

蜂胶对变形链球菌抑菌活性的研究进展

龋病是一种微生物感染性疾病,变形链球菌是引起其发生发展的主要致龋菌之一。近年来天然药物对龋病防治的研究已成为热点,而蜂胶是一种天然抗菌剂,国内外相关研究表明蜂胶对变形链球菌的生长、产酸、粘附、产胞外多糖及牙菌斑等方面有抑制的作用。本研究就蜂胶对变形链球菌的主要致龋毒力因子的作用研究作一综述。     

致龋菌与牙周致病菌混合感染动物模型的初步研究

目的通过建立龋病与牙周病共同发生的动物模型,初步探讨致龋菌与牙周致病菌的相关性。方法 5~6周龄金黄地鼠22只,雌雄各半,随机分成2组,均给予高蔗糖饮食。对照组10只,感染组12只,双侧下颌第一磨牙丝线结扎,接种牙龈卟啉单胞菌(P.g)和变形链球菌(S.m);种菌结束后8周检查下颌第一磨牙牙体和牙周组织损害情况。结果感染组产生明显的牙周损害:探诊出血(BOP)、菌斑指数(PLI)以及牙槽骨吸收(ABL)均显著大于对照组(P0.01),同时,感染组冠部龋坏率达到90.0%,大于对照组(66.7%),但差异没有统计学意义。2个组均未产生根龋。结论牙龈卟啉单胞菌与变形链球菌分别作用于牙周和牙体组织发生龋及牙周损害,二者没有显著的相互抑制作用,可在地鼠口腔中同时出现龋和牙周炎病损的模型。     

Isolation and characterization of the mutans streptococci from the dental plaques in Koreans

Mutans streptococci have been implicated as cariogenic bacteria in dental caries because they can produce high levels of dental caries-causing lactic acid and extracellular polysaccharide. The aim of this study was to isolate and characterize the mutans streptococci from the dental plaque obtained from Koreans. The dental plaque samples were collected from the anterior and molar teeth of both jaws in 155 subjects (aged 2 to 33.2 years, average age 13.7+/-4.7 years). The samples were diluted by 100-fold in 1x PBS and plated on mitis-salivarius bacitracin (MSB) agar plates. The mutans streptococci grown on MSB plates were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) targeting dextranase gene (dex). The mutans streptococci were identified at the species level using a 16S rDNA sequencing comparison method. The biochemical tests were carried out to biotype the mutans streptococci. Ninety-five strains of the mutans streptococci out of 358 colonies, which were derived from 141 subjects, were isolated. Of them, 77 strains and 18 strains were Streptococcus mutans and Streptococcus sobrinus, respectively. The biotyping data showed that 62, 1, 20, 10, and 2 strains were biotypes I, II, IV, V and variant, respectively. Of the two strains of variant biotype, one strains was similar to biotype IV except that it was positive to the arginine hydrolysis test. We considered this one strain a new biotype, and classified it as biotype VII. In conclusion, S. mutans and its biotype I was most frequently isolated in Korean dental plaque. The mutans streptococci strains isolated in this study might be useful for the study of the pathogenesis and the prevention of dental caries.     

龋坏引起的慢性牙髓炎牙髓组织中的细菌超微观察

对１０例龋坏引起的慢性牙髓炎、未穿髓的新鲜离体患牙,进行分段定位、半薄切片、革兰氏染色,光镜观察定位后,再行超薄切片,醋酸铀－柠檬酸铅复染,电镜进行超微结构观察。结果：（１）患牙牙髓组织中有５例观察到多种类型微生物存在,为球杆菌、球菌、杆菌。（２）牙髓组织中的细菌位于细胞内及细胞外,并处于各种状态,有生长繁殖阶段的也有固缩溶解阶段的。（３）深龋引起的长期有症状的慢性牙髓炎牙髓组织中有大量活细菌存在,细菌均存在于冠髓内,接近龋坏部位,周围及深层组织中未见菌体。并观察到组织细胞吞噬大量细菌后,细胞变性,出现核固缩、核仁消失,细胞膜不清楚。细胞周围有大量溶酶体出现。结果提示：深龋引起的长期有症状的慢性牙髓炎,尽管未穿髓,牙髓组织中,仍可有大量活细菌长期存在。     

Detection of DNA from periodontal pathogenic bacteria in biofilm obtained from waterlines in dental units

Direct person-to-person transmission of periodontal bacteria through saliva has recently been widely reported and dental units have been demonstrated to retract saliva from patients under treatment and to release it into the mouths of subjects undergoing the next operation. In this study the presence of a group of periodontal pathogenic bacteria inside waterlines in dental units was investigated using polymerase chain reaction (PCR) based methods. Briefly, 18 dental units of three different manufacturers were studied. Dental units were divided into two groups according to their prevalent use in routine practice. The first group consisted of nine dental units characterized by the frequent use of high-speed dental hand-pieces directly inside the mouth and in contact with patients' saliva. The second group, as a control, consisted of nine dental units where high-speed dental hand-pieces were not in use. A one cm section of the waterline tubing connected to the high-speed hand-piece was removed from each dental unit to evaluate the presence of DNA of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Treponema denticola. Two specimens were positive for Prevotella intermedia DNA. All the positive results were from samples obtained from dental units categorised in the first group. These findings clearly suggest that dental units have the potential to transmit periodontal pathogens. Manufacturers should be invited to design dental units that incorporate automated devices to disinfect DUWLs between patients with minimal effort by dental staff.