维吾尔族慢性牙周炎中牙龈卟啉单胞菌菌毛fimA毒力基因型的分布

目的:分析维吾尔族慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌菌毛fimA毒力基因型的分布情况.方法:收集52例维吾尔族慢性牙周炎患者的龈下菌斑,采用16S rRNA PCR法检测牙龈卟啉单胞菌,并根据菌毛fimA毒力基因型的特异引物,用聚合酶链反应(PCR)检测Ⅱ型fimA和Ⅳ型fimA菌株的分布.结果:16S rRNA PCR法检测牙龈卟啉单胞菌在龈下菌斑中阳性检出率是76.9%.牙周袋PPD＞6 mm位点龈下菌斑标本的P.gingivalis检出率高于4＜PPD≤6 mm采样的位点,2组差异有统计学意义(P＜0.05).牙龈卟啉单胞菌菌毛fimA毒力基因型在牙龈卟啉单胞菌感染者的检出率分别是:ⅡfimA型为37.5%,ⅣfimA型为22.5%.结论:维吾尔族慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌有较高的检出率.牙龈卟啉单胞菌存在fimA毒力基因多态性.     

维吾尔族慢性牙周炎中牙龈卟啉单胞菌菌fimA毒力基因型的分布

目的：分析维吾尔族慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌菌毛fimA毒力基因型的分布情况。方法：收集52例维吾尔族慢性牙周炎患者的龈下菌斑，采用16SrRNA PCR法检测牙龈卟啉单胞菌，并根据菌毛fima毒力基因型的特异引物，用聚合酶链反应（PCR）检测Ⅱ型fimA和Ⅳ型fima菌株的分布。结果：16SrRNA PCR法检测牙龈卟啉单胞菌在龈下菌斑中阳性检出率是76．9％。牙周袋PPD〉6mm位点龈下菌斑标本的P．gingivalis检出率高于4〈PPD≤6mm采样的位点，2组差异有统计学意义（P〈0．05）。牙龈卟啉单胞菌菌毛．fimA毒力基因型在牙龈卟啉单胞菌感染者的检出率分别是：Ⅱ fimA型为37．5％，ⅣfimA型为22．5％。结论：维吾尔族慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌有较高的检出率。牙龈卟啉单胞菌存在fima毒力基因多态性。     

成人牙周健康状况与fimA基因型牙龈卟啉单胞菌的相关性

目的分析不同fimA基因型牙龈卟啉单胞菌（P.gingivalis）在牙周健康人群和慢性牙周炎人群中的分布,探讨不同fimA基因型P.gingivalis与成人牙周状况的相关关系。方法收集牙周健康组（136例）和慢性牙周炎组（115例）的龈下菌斑样本,采用16S rRNA PCR法检测P.gingivalis,并根据各fimA基因型（Ⅰ～Ⅴ和Ⅰb）的特异性引物检测不同fimA基因型P.gingivalis菌株的分布,计算OR值和95%可信区间。结果牙周健康组和慢性牙周炎组龈下菌斑样本中P.gingivalis阳性率分别为22.1%和81.7%,多数样本中只检测到1种fimA基因型。牙周健康组中ⅠfimA型的检出率最高（占66.7%）;慢性牙周炎组中则为ⅡfimA基因型（占43.6%）,其次为Ⅳ和Ⅰb fimA基因型。慢性牙周炎的发生与P.gingivalis的关系密切（OR=16.36）,Ⅰ、Ⅰb、Ⅱ、Ⅲ、Ⅳ、ⅤfimA基因型P.gingivalis与慢性牙周炎相关性的OR值分别为0.97、13.26、36.62、4.57、22.86、1.19;ⅡfimA基因型P.gingivalis与慢性牙周炎的相关性最强,其次为Ⅳ和Ⅰb型。结论P.gingivalis菌株的fimA基因型存在差异,特异性fimA基因型P.gingivalis可能与成人慢性牙周炎的发生关系密切。     

不同fimA基因型牙龈卟啉单胞菌在青春期龈炎患者中的分布

目的:分析不同fimA基因型牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)在青春期龈炎患者和青春期牙周健康者中的分布。方法:收集51例青春期龈炎患者和46例青春期牙周健康者的龈下菌斑,采用16SrRNA PCR检测P.gingivalis,并根据各fimA基因型的特异引物,用PCR检测不同fimA基因型菌株的分布。结果:龈炎组各fimA基因型P.gingivalis的总检出率:Ⅰ型34.2%,Ⅱ型55.3%,Ⅳ型18.4%,Ⅲ型和Ⅴ型未检出;另有7例(占18.4%)未分出型别。健康组各fimA基因型P.gingivalis的总检出率:Ⅰ型14.3%,Ⅱ型85.7%,Ⅲ型14.3%,Ⅳ型28.6%,Ⅴ型未检出;另有2例(占14.3%)未分出型别。结论:青春期龈炎和青春期牙周健康者龈下菌斑中的P.gingivalis存在fimA基因多态性。fimA基因Ⅱ型是其主要存在的基因型,其次是Ⅰ型和Ⅳ型,Ⅲ型和Ⅴ型较少或不能检出。     

牙龈卟啉单胞菌菌毛fimA基因型在牙周牙髓联合病变中的分布

目的 分析牙龈卟啉单胞菌菌毛不同fimA基因型在牙周源性牙周牙髓联合病变患牙根管内的分布状况.方法 收集因牙周损害导致的牙周牙髓联合病变患牙43例,开髓揭髓顶后用纸尖采集根管内组织液,提取DNA.采用16SrRNA PCR检测牙龈卟啉单胞菌,并根据各fimA基因型的特异性引物,用聚合酶链反应检测不同fimA基因型菌株的分布.结果 16S rRNA PCR检测牙龈卟啉单胞菌阳性率76.7％.牙龈卟啉单胞菌fimA各基因型在牙龈卟啉单胞菌携带者的检出率分别为:Type Ⅰ fimA为24.2％;Type Ⅰ b fimA为27.2％;Type ⅡfimA为75.8％;Type ⅢfimA为9.1％;TypeⅣfimA为48.5％;Type ⅤfimA未检出;同时检测出Type Ⅰ和Ⅰb fimA为3.0％;Type Ⅱ和ⅣfimA为39.4％;Type Ⅰ、Ⅰb和ⅡfimA为21.2％.Type Ⅱ和Ⅳ fimA基因型的检出率显著高于其他基因型(P＜0.05).结论 牙周源性牙周牙髓联合病变根管内牙龈卟啉单胞菌存在fimA基因多态性.TypeⅡ和ⅣfimA基因型牙龈卟啉单胞菌菌株是牙周牙髓联合病变根管内的主要定植菌,可能与牙周牙髓联合病变的发生、发展相关密切.     

不同fimA基因型牙龈卟啉单胞菌在牙周健康人群中的分布

目的调查不同fimA基因型牙龈卟啉单胞菌 (P.gingivalis)在牙周健康人群中的分布情况。方法收集136例牙周健康者的龈下菌斑样本,采用16S rRNAPCR法检测P.gingivalis;并根据各fimA基因型特异性引物,用PCR法检测不同fimA基因型P.gingivalis的分布。 结果136例牙周健康者的龈下菌斑样本中携带P.gingivalis的阳性率为22.1%。大多数受检者龈下菌斑中只检测到1种fimA基因型 P.gingivalis菌株(80.0%);5例样本检出了2种fimA型P.gingivalis(16.7%),且均为ⅠfimA型与其它fimA 型P.gingivalis的联合检出。各fimA型P.gingivalis的检出情况:Ⅰ型(66.7%)、Ⅰb型(6.7%)、Ⅱ型(6.7%)、 Ⅲ型(10%)、Ⅳ型(6.7%)、Ⅴ型(16.7%)。Ⅰ型的检出率明显高于其它各型,差异有统计学意义(P<0.05);其它各fimA型 P.gingi-valis间的检出差异均无统计学意义。结论本研究条件下ⅠfimA型P.gingivalis在牙周健康人群中检出例数最高,提示:我 国牙周状况不同人群携带的P.gingivalis菌株fimA基因型可能存在差异。?     

不同rag基因型牙龈卟啉单胞菌在慢性牙周炎患者中的分布

目的分析不同rag基因型牙龈卟啉单胞菌在慢性牙周炎患者中的分布状况。方法收集50例慢性牙周炎患者的150个龈下菌斑样本,采用16S rDNA 聚合酶链反应（PCR）法检测牙龈卟啉单胞菌在牙周炎病变位点的检出率,并根据各rag基因型的特异性引物检测牙龈卟啉单胞菌不同rag基因型在慢性牙周炎病变位点的分布。结果经16S rDNA PCR法检测,病变位点牙龈卟啉单胞菌阳性检出率为70.7%。各rag基因型在牙龈卟啉单胞菌阳性位点的总检出率：rag-1为60.4%,rag-2为23.6%,rag-3为44.3%,rag-4为15.1%;经统计学检验,rag-1和rag-3型检出率较高,高于rag-2和rag-4型（P<0.05）。结论慢性牙周炎患者龈下菌斑中的牙龈卟啉单胞菌存在rag基因多态性,rag-1和rag-3基因型牙龈卟啉单胞菌与中国辽宁地区人群慢性牙周炎的发生发展关系密切。     

感染根管中牙龈卟啉单胞菌fimA基因多态性研究

目的研究根尖周炎患牙感染根管内牙龈卟啉单胞菌(P.gingivalis)不同fimA基因型的分布情况。方法收集200例感染根管内样本,经DNA抽提后使用16S r DNA PCR方法检测P.gingivalis。在P.gingivalis检出阳性样本中,根据各fimA基因型的特异引物,采用PCR检测不同fimA基因型菌株的分布,并通过Pearsonχ2检验对不同fimA基因型与临床症状进行相关性分析。结果感染根管中P.gingivalis的检出率为48%,其中79例样本只检测到1种fimA基因型P.gingivalis菌株(82.3%);12例样本检测到2种或2种以上fimA基因型P.gingivalis(12.5%)。各fimA基因型P.gingivalis的检出情况:Ⅰ型29.2%、Ⅰb型8.3%、Ⅱ型37.5%、Ⅲ型14.6%、Ⅳ型19.8%。各型fimA基因与临床症状之间无统计学相关性(P<0.05)。结论感染根管内P.gingivalis的fimA基因存在基因多态性,其中以fimAⅡ型P.gingivalis菌株检出率最高,其次为Ⅰ型和Ⅳ型。     

龈下菌斑中牙龈卟啉单胞菌牙龈素基因片段的检测

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,P.gingivalis）的2个基因片段kgp-cd和rgpB-cd,探讨2个基因片段的存在和缺失与牙周临床指标之间的关系.方法：选择慢性牙周炎患者龈下菌斑84个,对P.gingivalis阳性的龈下菌斑样本进行kgp-cd和rgpB-cd基因片段检测;根据2个基因片段的有无,将P.gingivalis分为A型和B型,采用SPSS11.5统计软件包,用t检验和x2检验分析不同基因型P.gingivalis与牙周临床指标的关系.结果：A型P.gingivalis在慢性牙周炎中的检出率高于B型（P＜0.05）,分别为85.29%和14.71%.不同基因型P.gingivalis引起的牙周袋探诊深度和牙龈出血倾向存在显著性差异（PD：t=2.85,P＜0.05;BOP：P＜0.05）.结论：kgp-cd和rgpB-cd基因与P.gingivalis的致病性有关.     

牙龈卟啉单胞菌在龈下菌斑和颊黏膜中的检测

目的　检测牙周健康者及牙周炎患者在颊黏膜和龈下菌斑中牙龈卟啉单胞菌的阳性率,探讨其与牙周炎发生和发展的关系。方法　选取40例牙周健康者和39例慢性牙周炎患者,分别收集颊黏膜和龈下菌斑样本,提取细菌DNA,设计细菌通用引物和牙龈卟啉单胞菌的特异引物用于PCR扩增,检测牙龈卟啉单胞菌的阳性率。结果　牙周健康组菌斑样本和颊黏膜样本牙龈卟啉单胞菌的阳性率分别为37·5%和32·5%,而牙周炎组菌斑样本和颊黏膜样本牙龈卟啉单胞菌的阳性率分别为69·23%和46·15%。牙周炎组菌斑的牙龈卟啉单胞菌阳性率高于牙周健康组,颊黏膜的牙龈卟啉单胞菌阳性率在组间无统计学差异;牙周炎组菌斑牙龈卟啉单胞菌阳性率高于颊黏膜, 牙周健康组两部位阳性率无统计学差异。结论　牙龈卟啉单胞菌除在菌斑中有高检出率外,在颊黏膜中也有较高的检出率,提示颊黏膜也是牙周细菌在口腔定植的重要部位,牙龈卟啉单胞菌也可在健康人群中检出,提示其有可能是口腔内固有菌群之一。     

Quantification of Porphyromonas gingivalis and fimA genotypes in smoker chronic periodontitis

Aim: Porphyromonas gingivalis fim A genotypes were associated with virulence factors in vitro , but little evidence of an association with disease severity were shown in humans. We aimed to correlate levels of P. gingivalis fimA genotypes II and IV and probing depth in smoker-chronic periodontitis subjects.
Material and Methods: One hundred and sixty eight subgingival samples of 20 smokers non-treated chronic periodontitis subjects obtained from sites with different probing depths [shallow (3 mm), intermediate (4–6 mm), deep (7 mm)] were analysed by real-time PCR for P. gingivalis and genotypes fimA II and IV.
Results: P. gingivalis and fimA IV were detected in all subjects, whereas fimA II was detected in 18 subjects (90%). One hundred and fifty two sites (90.5%) harboured P. gingivalis . Genotypes II and IV were detected in 28% and 69.6% of sites, respectively. The proportions of genotypes II and IV in relation to P. gingivalis levels were similar in shallow, intermediate and deep probing sites (2.4%, 4.6%, 1.4% for genotype II and 15.5%, 17.7%, 11.7% for genotype IV, respectively), indicating that other non-tested genotypes were more abundant. Increased levels of genotype IV were associated with increasing probing depth, but not of genotype II.
Conclusions: The data suggested an association between P. gingivalis genotype fimA IV and disease severity in smoker-chronic periodontitis subjects.     

Prevalence of fimA genotypes of Porphyromonas gingivalis and other periodontal bacteria in a Spanish population with chronic periodontitis

Objectives: The aim of this study was to determine the prevalence of the different fimA genotypes of Porphyromonas gingivalis in adult Spanish patients with chronic periodontitis, patients with gingivitis and periodontally healthy subjects, and the relationship between these genotypes and other periodontopathogenic bacteria. Study design: Samples of subgingival plaque were taken from 86 patients (33 with chronic periodontitis, 16 with gingivitis, and 37 periodontally healthy) in the course of a full periodontal examination. PCR was employed to determine the presence of the 6 fimA genotypes of Porphyromonas gingivalis (I-V and Ib) and of Aggregatibacter actinomycetemcomitans, Tannerella forsythia and Treponema denticola. Results: Porphyromonas gingivalis fimA genotypes II and Ib were present in significantly higher percentages in periodontal patients (39.4% and 12.1% respectively) than in healthy or gingivitis subjects. The prevalence of Tannerella forsythia, Treponema denticola, and Porphyromonas gingivalis fimA genotype IV was significantly higher in the group that presented bleeding greater than 30%. A positive correlation was found between Porphyromonas gingivalis fimA genotype IV and Treponema denticola. Conclusions: A strong association between Porphyromonas gingivalis fimA genotypes II and Ib and chronic periodontitis exists in the Spanish population. The most prevalent genotype in periodontal patients is II. Key words:Periodontitis, Porphyromonas gingivalis, fimA genotype, periodontal bacteria, polymerase chain reaction.     

Prevalence of Porphyromonas gingivalis fimA genotypes in Caucasians

The aim of the present study was to determine the prevalence of Porphyromonas gingivalis fimA genotypes in Caucasian patients with periodontitis. A total of 102 patients harboring P. gingivalis subgingivally were enrolled into the study. Pooled subgingival plaque samples of the six most severely affected sites were taken and analysed by fimA-specific polymerase chain reaction (PCR) and restriction analysis. Moreover, 26 P. gingivalis isolates were analysed by sequence analysis of the fimA gene. Sequence analysis revealed five major fimA genotypes (fimA types I-V) and allowed further subtyping of fimA genotypes II and IV into two subgroups each. The overall prevalences of fimA genotypes as assessed by PCR and restriction analysis among the P. gingivalis-positive patients with periodontitis were: type I, 25.5%; type II, 38.2%; type III, 4.9%; type IV, 18.6%; type V, 3.9%; and non-typable, 6.9%. Two patients were colonized by both type II and type IV, or type III and type IV fimA genotypes, respectively. Patients harboring different fimA genotypes showed no significant difference in severity of periodontal disease, as assessed by pocket probing depth and bleeding on probing following adjustment for smoking habit and age. The results indicate that predominant fimA genotypes in Caucasian periodontitis patients are types I, II, and IV. However, there was no difference in the association of the various fimA genotypes with disease severity.     

The prevalence and pathogenic differences of Porphyromonas gingivalis fimA genotypes in patients with aggressive periodontitis

BACKGROUND: The fimA gene, which encodes fimbrillin (FimA), is found in Porphyromonas gingivalis and has been classified into six genotypes based on nucleotide sequence. P. gingivalis that possesses the type II fimA gene is prevalent in adult periodontitis. OBJECTIVES: The aim of this study was to investigate the prevalence of P. gingivalis fimA genotypes in Japanese aggressive periodontitis patients, and to examine their virulence. METHODS: Subgingival plaque samples were obtained from 223 sites in 18 aggressive periodontitis patients and 95 sites in 22 periodontally healthy young adults. Actinobacillus actinomycetemcomitans, P. gingivalis and Tannerella forsythensis detection, determination of the fimA genotype in P. gingivalis, and the quantification of P. gingivalis were analyzed by polymerase chain reaction (PCR) methods. The proteolytic activities of the P. gingivalis fimA type I and fimA type II were also examined. RESULTS: In aggressive periodontitis patients, the most prevalent fimA genotype was the type II (46.7%), followed by the type Ib and type I, whereas in healthy subjects, the type I fimA was the only genotype detected. The number of P. gingivalis pathogens was the greatest in the type I fimA positive sites, and the frequency of coexisting A. actinomycetemcomitans and T. forsythensis was highest in the type II fimA positive sites in the aggressive periodontitis patients. Both the arginine-specific cysteine proteinase (Arg-gingipain) and lysine-specific cysteine proteinase (Lys-gingipain) activity of the P. gingivalis fimA type I strain were significantly higher than those of the fimA type II strains. CONCLUSIONS: These results suggest that differences in virulence exist among different fimA genotypes. Coadherence with other pathogens in P. gingivalis fimA type II-associated aggressive periodontitis and quantitative increases in P. gingivalis in fimA type I-associated aggressive periodontitis are related to this virulence.     

Relationship between transmission of Porphyromonas gingivalis and fimA type in spouses

BACKGROUND: Porphyromonas gingivalis is one of the major microbial pathogens associated with chronic periodontitis. To eradicate such pathogens by periodontal therapy, it is essential to clarify the source of infection. Recent findings suggest that the genotype of the fimbriae is one of the important factors in infection by P. gingivalis. The objectives of the present study were to investigate the transmission of P. gingivalis between spouses and to determine the relationship between P. gingivalis fimA type and colonization. METHODS: A total of 14 couples were selected to investigate the transmission of P. gingivalis and its association with the fimA types. To examine the distribution of fimA type in the general population, 32 subgingival plaque samples from 47 patients with periodontitis were also tested. The transmission of P. gingivalis strains was determined by using pulsed field gel electrophoresis (PFGE). P. gingivalis strains isolated from the couples and subgingival dental plaque samples were studied for fimA classification. RESULTS: The PFGE patterns of P. gingivalis strains from matched husbands and wives were identical for six of the 14 couples. In five of these six couples (83.3%), P. gingivalis strains harboring the type II fimA gene were present. The proportion of type II fimA in the strains isolated from couples with probable intrafamilial transmission was significantly higher than that in patients with periodontitis or in the group of samples isolated from one member of a couple. CONCLUSION: This study suggests that fimA type II, even though widely distributed in patients with periodontitis, may be an important factor in the transmission of P. gingivalis between spouses.     

慢性牙周炎患者龈下菌斑中不同基因型牙龈卟啉单胞菌的检测

摘要 目的:建立临床标本中牙龈卟啉单胞菌(P.g)的PCR检测方法,探讨慢性牙周炎患者不同牙位的龈下菌斑中P.g基因型的差异。方法:采用培养法分离鉴定慢性牙周炎患者不同牙位龈下菌斑中P.g,同时采用PCR检测 P.g16SrDNA、prtC和fimA基因。部分扩增产物测定了核苷酸序列。结果:在66例患者的127个龈下菌斑标本中, P.g16SrDNA、prtC和fimA多重引物扩增的阳性率为9814%;PCR阳性率显著高于培养法P.g的检出率(P< 0101)。3010%的患者(18/60)同时感染了不同基因型的P.g菌株。P.g16SrDNA、prtC和fimA扩增片段的核苷酸序列同源性在98162%~100%之间。结论:本文所建立的P.g的PCR检测方法具有较高的敏感性和特异性,适用于P.g的快速临床诊断。同一患者可被不同感染来源的多株P.g同时感染。     

Prevalence of fimA genotypes of Porphyromonas gingivalis and periodontal health status in Chinese adults

Background and Objective:  Porphyromonas gingivalis fimbriae play a key role in colonization of the oral cavity. The fimA gene, which encodes fimbrillin ( FimA ), can be classified into six types (I–V and Ib) according to nucleotide sequence. In the present study, we investigated the relationship between the prevalence of P. gingivalis -specific fimA genotypes and periodontal health status in Chinese adults.
Material and Methods:  One-hundred and fifteen patients with chronic periodontitis and 136 periodontally healthy adults were selected. P. gingivalis detection, determination of fimA genotypes, and the co-existence of Actinobacillus actinomycetemcomitans and Tannerella forsythia with various fimA types, were assessed by the polymerase chain reaction. Odds ratios and 95% confidence intervals were calculated for associating the fimA -specific genes with periodontitis.
Results:  P. gingivalis was detected in 22.1% of healthy subjects and in 81.7% of the patients. A single fimA genotype was detected in most samples. In healthy adults, the most prevalent fimA genotype was type I (66.7%). However, type II was detected most frequently (43.6%) in the patient group, followed by type IV (30.9%). The frequency of co-existing A. actinomycetemcomitans and T. forsythia was highest in type II fimA -positive sites. Statistical analysis revealed that periodontitis was associated with occurrences of type I (odds ratio 0.97), Ib (odds ratio 13.26), II (odds ratio 36.62), III (odds ratio 4.57), IV (odds ratio 22.86) and V (odds ratio 1.19).
Conclusion:  P. gingivalis type II followed by type IV were considered as disease-associated strains that account for the pathogenesis of chronic periodontitis in Chinese adults.