猛性龋儿童变链菌分离株的蔗糖粘附能力

目的探讨猛性龋儿童变链茵和远缘链球菌临床分离株的蔗糖依赖性粘附能力.方法采用紫外分光光度计,检测猛性龋、非猛性龋、无龋儿童变链茵(各6株)和远缘链球菌(猛性龋儿童6株,非猛性龋和无龋儿童各3株)临床株在含蔗糖培养液中对玻壁的粘附情况.结果各组变链茵分离株之间及各组远缘链球菌分离株之间对玻壁的粘附比无显著差异;各组远缘链球菌菌株对玻壁的粘附能力显著大于各组变链菌菌株(P＜0.05).结论猛性龋儿童变链菌和远缘链球菌临床株蔗糖依赖性粘附能力与非猛性龋及无龋儿童分离菌株无差别;远缘链球菌蔗糖依赖性粘附能力强于变链茵.     

猛性龋儿童变链菌分离株耐酸性实验研究

目的 :确定猛性龋儿童变链菌和远缘链球菌临床株的耐酸性。方法 :采用紫外分光光度计比较猛性龋、非猛性龋、无龋儿童变链菌 (各 6株 )和远缘链球菌 (猛性龋儿童 6株 ,非猛性龋和无龋儿童各 3株 )临床株在体外不同初始 pH条件下的生长情况。 结果 :初始 pH 4.5～ 5 .5条件下 ,各组变链菌生长抑制程度均明显大于远缘链球菌(P <0 .0 5 )。初始pH 4.5条件下 ,猛性龋儿童远缘链球菌分离株耐酸性明显强于非猛性龋和无龋儿童分离菌株(P <0 .0 5 )。结论 :远缘链球菌的耐酸性强于变链菌 ;猛性龋儿童远缘链球菌分离株耐酸性强     

猛性龋儿童变链菌分离株耐酸性实验研究

目的：确定猛性龋儿童变链菌和远缘链球菌临床株的耐酸性。方法：采用紫外分光光度计比较猛性龋、非猛性龋、无龋儿童变链菌（各6株）和远缘链球菌（猛性龋儿童6株,非猛性龋和无龋儿童各3株）临床株在体外不同初始pH条件下的生长情况。结果：初始pH4．5－5．5条件下,各组变链菌生长抑制程度均明显大于远缘链球菌（P＜0．05）。初始pH4．5条件下,猛性龋儿童远缘链球菌分离株耐酸性明显强于非猛性龋和无龋儿童分离菌株（P＜0．05）。结论：远缘链球菌的耐酸性强于变链菌;猛性龋儿童远缘链球菌分离株耐酸性强。     

儿童猛性龋变链菌分离株的产酸性

目的：确定儿童猛性龋变链菌和远缘链球菌临床分离株的产酸性。方法：采用酸度计和自动气相色谱仪比较儿童猛性龋、非猛性龋、无龋变链菌（各6株）和远缘链球菌（儿童猛性龋6株,非猛性龋和无龋各3株）的临床株降低环境pH值的能力（ΔpH）和乳酸产量,并以此推测其致龋力。结果：远缘链球菌ΔpH及乳酸产量均高于变链菌,特别是在低pH水平下,二者差异更为显著（P＜0．05）。儿童猛性龋远缘链球菌分离株ΔpH和乳酸产量显著大于非猛性龋和无龋分离菌株（P＜0．05）。结论：远缘链球菌的产酸能力强于变链菌;儿童猛性龋远缘链球菌分离株较非猛性龋和无龋儿童分离菌株产酸性更强。     

幼儿猛性龋病原菌的分离鉴定

目的:确定幼儿猛性龋的优势病原菌,为其防治提供依据。方法:采用细菌分离培养、形态学、生理生化学和 DNAG+C mol%测定方法,对30名2~5岁猛性龋患儿牙菌斑菌丛进行分离鉴定,采样部位为上颌患龋乳切牙龋损部位及邻近健康釉质表面,对照组的非猛性龋和无龋儿童则采集上颌乳前牙唇面颈1/3处的菌斑。结果:猛性龋儿童龋损部位变链菌和远缘链球菌的检出率及两个采样部位菌斑标本中变链菌和远缘链球菌的检出水平均显著高于非猛性龋和无龋儿童(P<0105)。结论:变链菌和远缘链球菌为幼儿猛性龋的优势病原菌。关键词 幼儿猛性龋变链菌远缘链球菌。     

高龋及无龋儿童变形链球菌分离株的蔗糖粘附能力比较

目的:探讨高龋和无龋儿童变形链球菌(变链菌)临床分离株的蔗糖依赖性粘附能力。方法:选取本实验室从10例高龋儿童和10例无龋儿童(3￣5岁)牙菌斑内分离、鉴定所得的60株变链菌临床分离株进行研究。采用紫外分光光度计,检测来自高龋儿童(dmfs≥6)的39株和无龋儿童(dmfs=0)的21株变链菌在含糖培养基中对玻壁的粘附情况。采用SPSS12.0软件包进行单因素方差分析,比较高龋儿童和无龋儿童之间牙菌斑内变链菌分离株的玻壁粘附能力。结果:在含1%蔗糖的培养基中,高龋组变链菌分离株对玻壁的粘附比平均值为(55.49+26.16)%,无龋组变形链球菌分离株对玻壁的粘附比平均值为(27.01+18.39)%,2组间差异具有显著性,P<0.01。结论:在含蔗糖环境中,分离自高龋儿童牙菌斑的变链菌株对玻壁的粘附能力高于来自无龋儿童的变链菌分离株,提示变链菌临床分离株的粘附能力与其致龋力相关。     

儿童猛性龋病原菌母子传播研究

目的 :探讨幼儿猛性龋病原菌母子传播途径。方法 :随机选择可获得变形链球菌群 (MS)纯培养的 2～ 5岁猛性龋、非猛性龋及无龋儿童母子各 10对 ,进行DNA指纹实验 ,检测儿童牙菌斑和母亲唾液中MS菌株的遗传相似性 ,并检查母亲唾液MS水平。结果 :猛性龋儿童MS菌株基因型与母亲的一致性为 70 % ,与非猛性龋和无龋儿童(均为 6 0 % )无显著差异 ;但猛性龋儿童MS基因型数目显著多于非猛性龋和无龋儿童 (P <0 0 5 )。猛性龋儿童母亲唾液变链菌水平与非猛性龋和无龋儿童母亲相似 ,但唾液远缘链球菌水平及DMFT均数显著高于无龋儿童母亲。结论 :母亲是儿童MS的主要来源 ;幼儿猛性龋与母亲的DMFT计数及唾液远缘链球菌水平成正相关 ,母子传播对幼儿猛性龋的发生具有一定作用。     

不同浓度的葡萄糖对变形链球菌初始粘附能力的影响

目的 研究不同浓度的葡萄糖对变形链球菌初始粘附能力的影响,同时观察高龋组、无龋组变形链球菌初始粘附能力的差别。方法 选取高龋组、无龋组变形链球菌（血清 c型）临床分离株各10株及1株参考株UA159,用唾液包被的羟基磷灰石（SHA）模拟口腔中牙面情况。各菌株分别在含3H的0.2%、1.0%、5.0%葡萄糖的液体培养基中培养,配成菌悬液。菌液与SHA作用90 min,清洗、吸干,液体闪烁计数测量各样本中粘附于SHA的细菌的量。各变形链球菌菌株在不同浓度葡萄糖条件下对SHA的粘附能力以粘附量CPM值表示。结果 变形链球菌高龋组分离株初始粘附能力明显高于无龋组分离株（P<0.05）;同时变形链球菌在不同浓度的葡萄糖营养条件下其初始
粘附能力也有统计学上的差别（P<0.05）,5.0%葡萄糖组粘附力最强,0.2%葡萄糖组粘附力最低,1.0%葡萄糖组粘附力界于两者之间。结论 ①变形链球菌的初始粘附能力可能与龋病的发生有关;②葡萄糖可能与变形链球菌的初始粘附相关,在一定范围内,葡萄糖可能会促进变形链球菌的 初始粘附。     

维吾尔族不同龋敏感儿童变链菌临床分离株生物膜状态合成胞外多糖能力的比较

目的:比较维吾尔族高龋和无龋儿童变链菌临床分离株在生物膜状态下合成胞外多糖的能力,以推测其致龋能力的差异。方法:1)选取课题组前期分离鉴定的维吾尔族儿童变链菌临床株27株,其中高龋(dmft≥5)17株,无龋(dmft=0)10株。2)在0.8cm×0.8cm无菌盖玻片上形成变形链球菌生物膜标本。3)采用蒽酮法测定高龋组与无龋组生物膜状态下合成胞外多糖的量。结果:高龋组合成水溶性及水不溶性葡聚糖量平均为(0.3011±0.0398)g/L和(0.3711±0.0372)g/L,高于无龋组的(0.2067±0.0265)g/L和(0.3489±0.0537)g/L,差异有统计学意义(P<0.05)。高龋及无龋组变链菌临床菌株生物膜状态下合成水不溶性葡聚糖量为(0.3656±0.0459)g/L高于水溶性葡聚糖量(0.2539±0.0586)g/L,差异有统计学意义(P<0.05)。结论:维吾尔族不同龋敏感儿童变链菌临床分离株生物膜状态下合成胞外多糖能力有差异,推测与其致龋性差异有关。     

变链菌分离株的形态、生理生化和遗传学鉴定

目的：探讨形态学和生理生化反应与DNA碱基含量测定鉴定变链菌和元缘链球菌临床分离株的一致性。方法：采用形态学和生理生化试验鉴定变链菌和远缘链球菌临床分离株，采用高效反相液相色谱法测定细菌DNA G＋Cmol%以检验生化鉴定的准确性。结果:形态及生化鉴定为变链菌和远缘链球菌的临床分离株，其DNA碱基含量分别为37．52和45．19，各自位于变链菌和远缘链球菌36－38和44－46的G＋Cmol％参考范围。结论：变链菌和远缘链球菌的生化鉴定结果与DNA G＋Cmol％测定结果一致。通过形态及生理生化鉴定基本可以鉴定变链菌和远缘链球菌。     

小沟聚合物探针实时检测变形链球菌和远缘链球菌

目的　寻求一种快速检测变形链球菌(S.mutans)和远缘链球菌(S.sobrinus)的方法,研究变形链球菌群在儿童猛性龋患者口中的分布。方法　设计并合成针对S.mutans和S.sobrinus葡糖基转移酶基因(gtf)的特异性引物和小沟聚合物探针(MGB探针),对9株变形链球菌群参考菌株直接提取DNA及扩增纯培养后提取DNA分别进行检测,比较二者检测结果的异同。采集92例猛性龋儿童菌斑样本,用MGB探针进行实时检测。结果　采用MGB 探针可以特异性地鉴别S.mutans和S.sobrinus,直接检测和扩增纯培养后的定性检测结果完全一致,前者荧光出现的时间略迟。92例猛性龋患儿中S.mutans检出率为96.7%,S.sobrinus检出率为32.6%,所有检出S.sobrinus的菌斑样本均可检出S.mutans。结论　采用特异性MGB探针方法可以对菌斑中S.mutans和S.sobrinus进行实时检测, 提高检测效率。     

表兄链球菌的检测与儿童猛性龋的关系

目的：探讨表兄链球菌(Streptococcus sobrinus,S. sobrinus)与儿童猛性龋的关系。方法：根据前期郑州市区猛性龋调查结果,随机抽样选择3～5岁儿童66例,其中猛性龋、普通高龋及无龋组各22例。采用TYCSB培养基作变形链球菌(Streptococcus mutans, S. mutans )及S. sobrinus初步筛选,结合生理生化鉴定,并采用聚合酶链反应作最终鉴定。采用 SPSS10.0软件包对实验组与对照组S.mutans和S.sobrinus的检出率进行χ²检验,组间均数作t检验。结果： S. sobrinus在各组儿童牙菌斑中均不能脱离S. mutans而单独检出,猛性龋组S. mutans检出率高于高龋组,差异无显著性(P>0.05),而2组儿童S. sobrinus检出率的差异有显著性(P<0.05);猛性龋组与无龋组S. mutans检出率差异显著(P<0.05),S. sobrinus检出率差异显著(P< 0.01)。同时检出S. mutans和S. sobrinus的样本,其猛性龋的发生率及龋失补牙数、龋失补牙面数和平滑面龋均数与只能检出S. mutans及2种细菌均不能检出的样本的差异有显著性(P< 0.01)。结论：S. mutans与S. sobrinus是儿童猛性龋的主要致病菌,S. sobrinus与儿童猛性龋的发生有关。S. sobrinus对儿童猛性龋的发生、发展具有协同作用。     

远缘链球菌与乳牙龋病关系的研究

目的：探讨远缘链球菌与乳牙龋病的关系。方法：采有ＴＹＣＳＢ培养基作细菌筛选,运用分子遗传学手段即ＤＮＡ的Ｇ＋ Ｃ ｍｏｌ％ 测定作最终鉴定。结果：患龋儿童组与无龋儿童组变形链球菌检出率有显著差异（Ｐ＜０ ．０１）,远缘链球菌检出率有显著差异（Ｐ＜ ０．０５） ;患龋儿童组中能同时检出变形链球菌和远缘链球菌的群体,其龋失补牙数、龋失补牙面数及平滑面龋数均与只能检出变形链球菌的群体有显著差异（ Ｐ＜ ０．０１） 。结论：变形链球菌是龋病的主要致病菌,远缘链球菌对龋病过程有协同作用,与乳牙平滑面龋的发生、发展有关。     

Human strains of mutans streptococci show different cariogenic potential in the hamster model

The cariogenic potential of fresh humans strains of Streptococcus mutans and Streptococcus sobrinus was examined in hamsters maintained on a high sucrose diet. The strains 1B 16 (S. mutans) and B13 (S. sobrinus) isolated 20-25 yrs ago served as positive controls. Three series of experiments were run. Some strains were tested once and some strains were tested in all 3 experiments. The animals infected with S. sobrinus strains generally showed lower caries scores than those infected with S. mutans strains. Among the groups infected with the different strains of S. mutans the caries scores varied. In one of the experiments the caries score of the animals infected with one of the Icelandic S. mutans strains was significantly higher than that of the positive control group and the group infected with one of the fresh Swedish isolates. Thus, different strains of mutans streptococci where shown to induce varying caries activity in the hamster model.     

Mutans streptococci and lactobacilli in breast-fed children with rampant caries.

This study aimed to investigate the prevalence of selected components of the oral microflora in breast-fed children who developed rampant caries (resembling nursing caries) under hitherto unexplained circumstances. Dental plaque and saliva samples were collected from breast-fed children, aged between 1 and 2.5 years, with and without rampant caries. Mutans streptococci and lactobacilli were isolated from dental plaque of all children with rampant caries and from most caries-free children. None of the colonies of mutans streptococci resembled those of Streptococcus sobrinus. The mean counts of the mutans streptococci and lactobacilli were 100-fold higher in plaque samples from children with rampant caries as compared with caries-free children. No difference could be found between the numbers of mutans streptococci in plaque overlaying cavities and that from adjacent sound enamel. In contrast, the counts of lactobacilli in plaque were approximately 100-fold higher from cavities than from sound surfaces. The levels of mutans streptococci in saliva were directly related to the presence of rampant caries. The results show that caries-free and caries-active breast-fed children, aged 1 to 2.5 years, harbour mutans streptococci and lactobacilli on their teeth. Rampant caries in these children can occur in the absence of nursing bottles or any other feeding abuse during weaning and in the presence of an aciduric plaque microflora, as has been reported for children with nursing bottle caries.     

Differentiation of mutans streptococci by intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

It is difficult to distinguish mutans streptococci on the species level, and even more so on the subspecies level. Intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) (ICM) was applied to reference strains of five of the species of the mutans group (Streptococcus criceti, Streptococcus downei, Streptococcus mutans, Streptococcus ratti, Streptococcus sobrinus), nonmutans streptococci (Streptococcus oralis, Streptococcus mitis, Streptococcus salivarius, and Streptococcus sanguinis), and 177 mutans streptococci isolated from saliva of 10 children. From the analysis of the reference strains, readily distinguishable ICM mass spectra were obtained for the different species. Based on multivariate statistical analysis, a correct and unambiguous assignment was made of the spectra of 159 isolated mutans streptococci to S. mutans and 16 isolates to S. sobrinus. Two isolates were sorted out and were identified by sequencing of their 16S rRNA genes as Streptococcus anginosus. In addition, ICM indicated a misclassification for some reference strains (AHT, V 100 and E 49) and re-classified AHT and E 49 as S. ratti and V 100 as S. sobrinus. This was confirmed by 16S rDNA sequencing. Based on a statistical similarity analysis of the spectra of reference strains and a quantitative assessment of the reproducibility of ICM, the isolates identified as either S. mutans or S. sobrinus were phenotyped on the subspecies level. In the population of the clinical isolates, 14 unambiguously different S. mutans and three different S. sobrinus phenotypes were detected. ICM proved to be a powerful tool for a differentiation of mutans streptococci down to the subspecies level.