牙龈卟啉单胞菌fimA基因型研究进展

牙龈卟啉单胞菌是一种非酵解糖的革兰阴性厌氧球杆菌,是目前研究广泛且证据充足的重要牙周致病菌之一。其中菌毛(fimbriae)对牙龈卟啉单胞菌的毒力起到了举足轻重的作用,不同基因型的菌毛又会导致不同的临床症状。本文回顾近几年牙龈卟啉单胞菌fimA不同基因型在牙周、牙体牙髓以及致病性机理方向上的研究进展。     

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。     

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。?     

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.     

不同fimA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fimA基因型牙龈卟啉单胞菌(P.gingivalis)在慢性牙周炎患者中的分布状况。方法　收集101例慢性牙周炎患者的龈下菌斑,采用常规培养法和16S rRNA PCR检测P.gingivalis,并根据各fimA基因型的特异引物,用聚合酶链反应(PCR)检测不同fimA基因型菌株的分布。结果　16S rRNA PCR检测P.gingivalis阳性检出率为88·1%。大多数受检牙龈下菌斑中只检测出一种fimA基因型菌株(65·1%),各fimA基因型的总检出率: ⅠfimA为24·7%;ⅡfimA为43·8%;ⅢfimA为15·7%;ⅣfimA为40·4%;VfimA为3·4%。结论　慢性牙周炎患者龈下菌斑中的牙龈卟啉单胞菌存在fimA基因多态性,ⅡfimA和ⅣfimA基因型P.gingivalis菌株与慢性牙周炎的发生发展关系密切。     

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。     

牙龈卟啉单胞菌研究进展

牙周炎是常见的口腔疾病之一,是中国成年人失牙的主要原因.牙龈卟啉单胞菌在引发牙周组织破坏过程中发挥着重要的作用,成为国内外学者研究的热点之一.本文就牙龈卟啉单胞菌与口腔疾病和其他全身性疾病的关系、牙龈卟啉单胞菌全基因组,牙龈卟啉单胞菌与牙龈上皮细胞的相互作用等研究作一综述,以拓宽牙龈卟啉单胞菌的研究思路,进血为牙周炎的...     

牙龈素在牙龈卟啉单胞菌生长及牙周致病过程中的作用

牙龈素是慢性牙周炎的重要致病菌---牙龈卟啉单胞菌的最重要毒力因子。作为一组半胱氨酸蛋白酶,牙龈素能结合并降解多种宿主蛋白质。本文总结了目前对牙龈素的结构和功能的研究,主要从牙龈素对牙龈卟啉单胞菌生长代谢、对牙周组织的致病作用及宿主免疫的影响三个方面阐述。     

两种fimA基因型牙龈卟啉单胞菌刺激下口腔上皮细胞ICAM-1的表达

目的比较2种fimA基因型牙龈卟啉单胞菌（Porphyromonas gingivalis,P．gingivalis）刺激下口腔上皮细胞ICAM-1的表达水平。方法实验组用P．gingivalis ATCC 33277（Ⅰ型菌毛组）,W83（Ⅳ型菌毛组）,47A-1（Ⅳ型菌毛组）分别与KB细胞（ATCC CCL17）共同孵育24h;对照组为未受P.gingivalis刺激的KB细胞。分别在1h、3h、6h和24h收集细胞,运用流式细胞仪检测KB细胞膜上ICAM-1的动态表达。结果P．gingivalis刺激细胞后3h、6h和24h,实验组ICAM-1的表达水平均高于对照组;2种fimA基因型P．gingivalis调节KB细胞表达ICAM-1的方式相似,Ⅳ型菌毛组的调节作用强于Ⅰ型菌毛组。结论P.gingivalis上调口腔上皮细胞表达ICAM-1的水平与其fimA基因型相关,提示P.gingivalis致病性与其fimA基因型相关。     

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。     

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.     

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。     

Comparison of inflammatory changes caused by Porphyromonas gingivalis with distinct fimA genotypes in a mouse abscess model

The fimA gene of Porphyromonas gingivalis, encoding fimbrillin (a subunit protein of fimbriae) has been classified into six genotypes (types I-V and Ib). The genotypic variation was previously suggested to be related to the severity of adult periodontitis in the general population. In this study, we compared inflammatory changes caused by bacterial infection to study pathogenic heterogeneity among the different fimA strains in a mouse abscess model. Bacterial suspensions of 13 P. gingivalis strains representing the six fimA types were subcutaneously injected into female BALB/c mice, and serum sialic acid concentrations were assayed as a quantitative host inflammatory parameter. Type II fimA organisms caused the most significant induction of serum sialic acid, as well as other infectious symptoms, followed by types Ib, IV and V. In contrast, types I and III caused weak inflammatory changes. In addition, fimA mutants of type II strains clearly lost their infectious ability. These findings suggest that fimA genotypic variation affects expression of P. gingivalis virulence.     

Identification of a new variant of fimA gene of Porphyromonas gingivalis and its distribution in adults and disabled populations with periodontitis

Porphyromonas gingivalis fimbriae are critical for the promotion of bacterial infection. The fimA gene encoding fimbrillin, a subunit of fimbriae, has been classified into five genotypes (types I to V) based on their nucleotide sequences. Using a fimA type-specific PCR assay, our previous study demonstrated a close relationship between P. gingivalis possessing type II and type IV fimA genes and adult periodontitis. In that study, some clinical specimens were found to be positive for both types I- and II- fimA specific primers, likely due to the coexistence of two clonal types or a single clone of an unknown genotype in the samples. In the present study, we cloned a new variant of the fimA gene, designated as type Ib fimA, from P. gingivalis HG1691. The nucleotide sequence of the cloned fimA gene showed a 97.1% homology with that of type I fimA, indicating it as a clonal variant of type I fimA. Organisms with type Ib fimA were detected in 13.5% of periodontitis patients and in 2.9% of periodontal healthy adults. Statistical analysis revealed a strong relationship between periodontitis and specific fimA types such as type Ib [odds ratio (OR) 6.51], type II (OR 77.8), and type IV (OR 7.54). Moreover, type Ib fimA-organisms were also found to be related to periodontitis in Down's syndrome (OR 1.91) and mentally disabled populations (OR 4.00). These findings suggest that P. gingivalis with type Ib fimA is closely associated with the progression of periodontitis, similar to organisms with type II and IV fimA.     

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.     

fimA genotypes and PFGE profile patterns in Porphyromonas gingivalis isolates from subjects with periodontitis

Background and objectives:  Porphyromonas gingivalis is frequently identified to type by evaluation of fimA polymorphisms and less often by pulsed-field gel electrophoresis (PFGE) because of the technical intricacies of PFGE. To compare these techniques, we genotyped P. gingivalis clinical isolates as to (i) their fimA type and (ii) their whole genome restriction profile (PFGE analysis).
Material and methods:  Thirty-two P. gingivalis strains were isolated from 16 unrelated periodontitis patients. Two strains were isolated from each patient. Strains were subjected to a fimA -typing polymerase chain reaction (PCR) assay. Strains that could not be typed by PCR were submitted to sequencing of the entire fimA gene. The PFGE profiles of clinical strains were compared using bioinformatic analysis.
Results:  Seven of the 32 isolates were not typeable by PCR and so their entire fimA gene was sequenced. The sequencing identified each strain as belonging to a single fimA type. In one case, sequencing of the fimA gene did not agree with the result obtained using fimA PCR typing. With the exception of one patient, each patient presented isolates bearing the same fimA type. However, in three patients, isolates with the same fimA type presented different PFGE pulsotypes.
Conclusion:  The P. gingivalis typing using fimA PCR has limitations in typeability and discriminatory power. A typing technique for P. gingivalis that is easy to perform but that presents adequate typeability and discriminatory power is needed if we want to better understand the epidemiology of periodontal disease.     

不同fimA基因型牙龈卟啉单胞菌刺激口腔上皮细胞白细胞介素-8的表达

目的比较不同fimA基因型牙龈卟啉单胞菌（P.gingivalis）刺激下口腔上皮细胞白细胞介素-8（IL-8）的表达水平,初步探讨P.gingivalis致病性与其fimA基因型的关系。方法P.gingivalis ATCC 33277（Ⅰ型fimA）、W83、47A-1（Ⅳ型fimA）和KB细胞ATCC CCL-17共同孵育24 h,以未受刺激的KB细胞作为对照组,分别在1、3、6、24 h收集细胞和培养上清液。RT-PCR检测KB细胞IL-8 mRNA的动态表达,酶联免疫反应检测培养上清液中IL-8的动态变化。结果2种fimA基因型菌株刺激1 h,KB细胞IL-8 mRNA的表达上调至峰值,高于对照组,3~24 h IL-8 mRNA的表达水平接近对照组;P.gingivalis感染细胞后3~24 h,上清液中的IL-8水平低于对照组,Ⅳ型菌株低于Ⅰ型菌株;IL-8 mRNA及其蛋白表达不完全一致,提示IL-8的表达存在转录后水平的调节。结论fimA基因型与口腔上皮细胞IL-8的表达水平相关,提示P.gingivalis致病性与其fimA基因型相关。