不同牙周状态下龈下菌斑中人类巨细胞病毒的检测

目的 :研究人类巨细胞病毒 (HCMV)与慢性牙周炎及其活动性的相关性。方法 :采集 62例慢性牙周炎患者 (男性 2 7例 ,女性 3 5例 )的牙周炎活动部位 ,牙周炎静止部位 ,以及轻度龈炎部位的龈下菌斑 ,使用SeekVi ralDNAkit试剂盒提取DNA ,采用巢式PCR法检测HCMV ,比较同一患者不同牙周状态的检出率并加以分析。结果 :慢性牙周炎患者牙周炎活动部位、牙周炎静止部位、轻度龈炎部位的HCMV检出率为分别为 3 8.7%、14 .5 %、12 .9% ;HCMV在牙周炎活动部位的检出率高于牙周炎静止期部位 (P <0 .0 5 ) ,以及轻度龈炎部位 (P <0 .0 5 ) ;牙周炎静止部位与轻度龈炎部位的HCMV检出率的差异无统计学意义 (P >0 .0 5 )。结论 :提示HCMV感染与慢性牙周炎及其活动性的有一定的相关性。     

慢性牙周炎与爱泼斯坦-巴尔病毒和人疱疹病毒6型的相关性

慢性牙周炎是一种最常见的多因素、炎症性和破坏性疾病,其发病与微生物、宿主和环境等多种因素有关,但其发病机制至今不明。研究显示,疱疹病毒与慢性牙周炎的发生发展有一定的相关性。爱泼斯坦-巴尔病毒(EBV)、人疱疹病毒(HHV)-6型均属疱疹病毒,本文就EBV、HHV-6以及二者间的关系,EBV和HHV-6引发慢性牙周炎的相关机制,EBV与慢性牙周炎,HHV-6与慢性牙周炎等研究进展作一综述。     

古细菌在牙周病患者龈下菌斑中的定性和定量分析

目的 探讨古细菌与牙周疾病的关系,为深入研究古细菌与人类疾病的相关性和古细菌的可能致病机制奠定基础.方法 收集侵袭性牙周炎23例、慢性牙周炎29例、慢性龈炎35例及牙周健康者38人的龈下菌斑,进行古细菌的定性和定量检测.结果 古细菌在龈下菌斑中的检出率:侵袭性牙周炎组65%,慢性牙周炎组72%,慢性龈炎组26%,健康对照组未检出.每微克湿重菌斑中的古细菌16S rRNA基因平均拷贝数:侵袭性牙周炎组6.66×106,慢性牙周炎组4.47×106,慢性龈炎组1.78×106.龈下菌斑中牙周炎组古细菌的检出率、古细菌16S rRNA基因平均拷贝数均高于龈炎组,差异有统计学意义(P＜0.05).结论 古细菌可能为牙周炎的致病因素之一.     

牙周机械治疗对慢性牙周炎患者龈沟液中疱疹病毒的影响

目的 通过纵向观察慢性牙周炎患者(chronic periodontitis,CP)治疗前及治疗后不同时间点龈沟液中两种疱疹病毒检出率的变化,分析疱疹病毒与牙周炎症的关系,为评价牙周炎治疗效果及判断预后提供一个微生物学手段.方法 应用巢式聚合酶链反应(nested PCR,n-PCR)技术,检测13例CP患者牙周机械治疗前、治疗后2周、2个月、4个月时龈沟液中的人巨细胞病毒(human cytomegalovirus,HCMV)、EB病毒1型(Epstein-Barr vires-1,EBV-1),并分析两种病毒检出率的变化.结果 CP患者在牙周机械治疗后2个月和4个月的菌斑指数、探诊深度和出血指数均较治疗前显著降低(P<0.01),但治疗后4个月的菌斑指数、探诊深度和出血指数均较治疗后2个月显著升高(P<0.05).CP患者治疗前HCMV、EBV的检出率分别为42%(33/78)、14%(11/78);EBV阳性位点多伴有HCMV感染,11个EBV(+)的位点中9个位点伴HCMV(+).HCMV、EBV检出的位点多是深牙周袋位点,14个深牙周袋位点中13个HCMV(+);11个EBV(+)位点中9个为深牙周袋位点.CP患者在治疗后2周时HCMV、EBV的检出率降至最低(分别为8%、0);治疗后2周、2个月、4个月HCMV和EBV的检出率均明显低于治疗前(P<0.01),HCMV在治疗后2个月的检出率(15%)较治疗后2周时(8%)回升,但差异无统计学意义(P=0.133).结论 疱疹病毒感染在CP发病中可能有一定作用;CP患者在牙周机械治疗后疱疹病毒的变化早于临床指标的变化,治疗后2个月内即应开始定期复查、复治.     

未获培养的微生物与牙周病关系初探

目的:研判未获培养微生物与牙周病的相关性。方法:针对16S rRNA基因设计未获培养微生物D084、I025特异探针,通过检测临床菌斑判断其与牙周病的关系。结果:引物特异性通过PCR产物测序获得验证,在慢性牙周炎组和单纯性龈炎组的D084检出率分别为62.9%和84.6%,I025为71.4%和42.3%,与正常组均有显著性差异(P<0.01)。I025在慢性牙周炎组和单纯性龈炎组间的检出率也存在显著性差异(P<0.05)。结论:未获培养微生物D084、I025与单纯性龈炎、慢性牙周炎患病相关,I025还与牙周炎进展相关。     

EB病毒和牙龈卟啉单胞菌协同感染与妊娠期慢性牙周炎的相关性研究

目的：研究妊娠期女性EB病毒（Epstein?Barr virus,EBV）和牙龈卟啉单胞菌（Porphyromonas gingivalis, Pg）的协同感染情况和牙周炎严重程度的关系。方法收集36例患慢性牙周炎（妊娠期慢性牙周炎组）的妊娠期女性和36例牙周健康（妊娠期牙周健康组）的妊娠期女性唾液样本,应用巢式PCR技术检测EBV感染率,应用16S rRNA为基础的PCR技术检测Pg感染率,结合口腔临床检查收集的牙周临床指标,对EBV和Pg的协同感染与牙周炎严重程度的相关性加以分析。结果妊娠期慢性牙周炎组和妊娠期牙周健康组EBV、Pg感染率的检出率差异无统计学意义（P＞0.05）,2组EBV和Pg协同感染的检出率差异具有统计学意义（χ2=4.800,P=0.028）。EBV和Pg协同感染与探诊出血指数相关（t=3.058,P=0.003）,与牙周袋深度和附着丧失无关（P＞0.05）。结论 EBV和Pg协同感染与妊娠期慢性牙周炎的相关性仍需进一步研究。     

EB病毒与逆行性牙髓炎的相关性研究

目的 研究EB病毒(EBV)与逆行性牙髓炎的相关性.方法 应用巢式聚合酶链反应(nPCR)技术,检测30例逆行性牙髓炎患者和30例健康对照者牙髓组织中的EBV.比较分析它们的检出率.结果 30例逆行性牙髓炎患者的EBV检出率为36.67%,显著高于健康对照者(0%),差异有统计学意义(P＜0.01).结论 提示EBV与逆行性牙髓炎有相关性.     

慢性牙周炎与爱泼斯坦-巴尔病毒和人疱疹病毒6型的相关性

慢性牙周炎是一种最常见的多因素、炎症性和破坏性疾病,其发病与微生物、宿主和环境等多种因素有关,但其发病机制至今不明。研究显示,疱疹病毒与慢性牙周炎的发生发展有一定的相关性。爱泼斯坦-巴尔病毒（EBV）、人疱疹病毒（HHV）-6型均属疱疹病毒,本文就EBV、HHV-6以及二者间的关系,EBV和HHV-6引发慢性牙周炎的相关机制,EBV与慢性牙周炎,HHV-6与慢性牙周炎等研究进展作一综述。     

不同程度慢性牙周炎患者病情的二年自然进展

目的　观察不同程度慢性牙周炎的自然进展规律。方法　纵向观察 16 9例轻、中、重度慢性牙周炎患者在 2年中的疾病自然进展 ,检查除第三磨牙外的全口牙 ,每颗牙 6个位点 ,以探诊深度、附着丧失为指标 ,两次检查之间附着丧失加重≥ 3mm的牙位定为活动性进展。结果　活动性发生率依基线时疾病的轻、中、重程度而逐渐增高 ,按位点活动性的发生率分别为 0 14 %、0 39%及0 73% ,按个体活动性的发生率分别为 15 5 6 %、2 9 89%及 4 3 2 4 % ,差异有统计学意义。结论　重度牙周炎患者是牙周破坏活动性进展的高危人群。     

不同引物检测慢性牙周炎龈下菌斑中齿垢密螺旋体

目的：利用两个不同引物对慢性牙周炎患者患病部位及相对健康部位龈下菌斑中齿垢密螺旋体（Td）进行检测,以了解Td在慢性牙周炎患者不同部位的分布及Td检出率与牙周炎临床指标的关系。方法：收集58例慢性牙周炎患者患病部位及相对健康部位龈下菌斑标本,利用PCR分别扩增53kDa外膜蛋白表达基因tdpA片段及16srRNA保守区片段。结果：58个患病部位龈下菌斑标本中tdpA及16srRNA扩增的阳性率分别为58．6％和81．0％,而相对健康部位龈下菌斑标本中PCR阳性率分别为8．62％及15．5％,患病部位Td检出率高于相对健康部位（P〈0．001）,16srRNA基因片段引物检出率高于tdpA基因片段（P〈0．05）。临床附着丧失≥5mm的患牙龈下菌斑标本中Td的检出率高于临床附着丧失〈5mm标本（P〈0．05）,不同牙周袋深度及牙龈指数标本的Td检出率之间差异无统计学意义（P〉0．05）。结论：在慢性牙周炎患者活动部位龈下菌斑中Td检出率高于相对健康部位;Td感染与慢性牙周炎关系密切;利用16srRNA保守区片段对齿垢密螺旋体进行检测检出率高于tdpA基因片段。     

牙周炎患者唾液中伴放线放线杆菌的检出状况分析

目的 检测不同类型牙周炎患者唾液中的伴放线放线杆菌(Actinobacillusactinomycetemcomitans,Aa),探讨唾液和集合龈下菌斑中Aa检出率的差异以及唾液中Aa的存在状况与牙周临床指标的关系. 方法 收集50例侵袭性牙周炎(aggressive periodontitis,AgP)患者、48例慢性牙周炎(chronic periedontitis,CP)患者和25例非牙周炎者的非刺激性全唾液和集合龈下菌斑,应用聚合酶链反应(PcR)技术检测两种样本中的Aa. 结果 Aa在AgP患者唾液中的检出率(32%)显著高于非牙周炎者(4%)和CP患者(15%),差异均有统计学意义(P<0.01,P<0.05),同时Aa在AgP患者唾液中的检出率也显著高于集合龈下菌斑样本(16%),差异亦有统计学意义(P<0.05).年龄≤30岁是唾液中存在Aa的危险指征(OR=3.23,P<0.05);出血指数≥3的位点超过70%与唾液中存在Aa有关(OR=19.21,P<0.01). 结论 AgP患者唾液样本中Aa的检出率明显高于集合龈下菌斑样本,亦高于CP患者和非牙周炎者,提示Aa可能参与AgP的发生和发展.     

Detection of herpetic viruses in gingival crevicular fluid of patients suffering from periodontal diseases: prevalence and effect of treatment

Background/Aim:  Although the role of bacteria in the etiology of periodontitis is well established, it has been suggested that herpetic viruses could contribute to the initiation and progression of this disease. The aim of this study was to determine the prevalence of human cytomegalovirus (HCMV), Epstein–Barr virus (EBV) and herpes simplex virus (HSV) in gingival crevicular fluid (GCF) samples obtained from periodontally healthy, gingivitis and periodontitis patients. In addition, the effect of periodontal treatment (scaling and root planing) on the persistence of herpetic viruses was evaluated in a sub-group of patients suffering from chronic periodontitis.
Methods:  The presence of viruses in GCF samples was assessed by a nested PCR amplification technique. The persistence of viruses in periodontal sites was evaluated following a scaling and root planing therapy.
Results:  A statistically significant higher prevalence of HCMV was observed in periodontitis patients as compared to healthy control subjects (35 vs. 8%, respectively; P  = 0.0377). A trend for a higher prevalence of HSV was also noted in the periodontitis group, in comparison with healthy control subjects. In addition, a higher prevalence of HCMV was associated with deep periodontal pockets in subjects suffering from periodontitis. In the sub-group of periodontitis patients, periodontal therapy resulted in the elimination (HCMV and EBV) or reduction (HSV) of the herpetic viruses.
Conclusions:  This study showed that the prevalence of HCMV and HSV viruses in GCF is higher in patients suffering from periodontitis compared to periodontally healthy subjects, and that the prevalence of HCMV is higher in deep periodontal pockets. It also brought evidences that periodontal therapy may be associated with virus elimination in diseased sites.     

Increased interleukin-18 in gingival crevicular fluid from periodontitis patients

INTRODUCTION: This study aimed to measure the levels of interleukin-18 (IL-18) in inflamed shallow sites and inflamed deep sites in patients with periodontitis and to compare the data with results from inflamed shallow sites in patients with gingivitis. A secondary aim was to examine the composition of the subgingival microbiota in the sampled sites. METHODS: Gingival crevicular fluid was collected from five gingivitis sites and five periodontitis sites from 18 patients with chronic periodontitis, and from five gingivitis sites from 15 patients with gingivitis. Samples from each site category were pooled and IL-18 levels were measured using an enzyme-linked immunosorbent assay. The subgingival microbiota was analyzed by checkerboard DNA-DNA hybridization. RESULTS: All clinical parameters and gingival crevicular fluid volumes were higher in periodontitis sites compared with gingivitis sites from patients with periodontitis and gingivitis. The total amount of IL-18 was higher in periodontitis sites than gingivitis sites in both periodontitis (P = 0.018) and gingivitis (P = 0.002) patients and was higher in gingivitis sites from periodontitis patients than in those from gingivitis patients (P = 0.015). There were higher levels of Tannerella forsythia, Porphyromonas gingivalis, and Treponema denticola (red complex species) in periodontitis sites compared with gingivitis sites in both the periodontitis and gingivitis patients (P < 0.001). CONCLUSION: Levels of IL-18 were higher in patients with chronic periodontitis compared with patients with gingivitis, even at sites with similar pocket depths. The presence of similar levels of red complex species in gingivitis sites from periodontitis patients and from gingivitis patients suggested that the higher levels of IL-18 were not associated with a different microbial challenge.     

Mammalian viruses in human periodontitis

A prior investigation has demonstrated a higher prevalence of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) in subgingival specimens from periodontitis patients than from gingivitis patients. This study aimed to determine the frequency of HCMV, EBV-1, EBV-2, herpes simplex virus (HSV) and human immunodeficiency virus (HIV) in subgingival samples from 27 adults who each contributed both a periodontitis and a gingivitis site. Viral detection was performed using a nested-polymerase chain reaction method. Twenty-four subjects (89%) yielded at least one of the five test viruses from deep periodontal pockets, wheras only 15 (56%) showed viruses from shallow periodontal sites ( P =0.015; chi-square test). Viral co-infection occurred more frequently in deep than in shallow periodontal sites ( P =0.015). HCMV was detected with higher frequency in deep than in shallow periodontal sites ( P =0.023). The possible periodontopathogenic mechanisms of mammalian viruses in human periodontitis are discussed. The role and importance of HCMV and other mammalian viruses in the initiation and progression of destructive periodontal disease merits further investigation.     

Crevicular fluid myeloperoxidase at healthy, gingivitis and periodontitis sites

The volume and myeloperoxidase (MPO) activity of gingival crevicular fluid (GCF) collected with filter paper strips for 30 s from the sulcus of healthy, gingivitis and periodontitis sites of Chinese subjects were measured. MPO/site and MPO/microliter GCF were both greater at gingivitis and periodontitis sites than at healthy sites. Enzyme activity was similar at the 2 categories of diseased sites. Mean GCF volume and MPO activity were calculated for all samples from healthy, gingivitis and periodontitis sites with GI 0, 1, 2 and 0 + 1. GCF volume, MPO/site and MPO/microliter GCF all were greater at GI 2 than GI 0 or 0 + 1. These data indicate that increased GCF MPO previously observed at periodontitis sites is not specific to such sites. Rather increased GCF MPO likely occurs when additional polymorphonuclear leukocytes enter the sulcus as a result of gingival inflammation. A second sample was obtained from 22 sites 4 weeks after the initial collection. These samples were collected for 5 s rather than 30 s. The GCF volume, MPO/site and MPO/microliters GCF were each greater in samples collected for 30 s rather than 5 s. Correlation coefficients showed that the amount of GCF and MPO activity of the fluid collected for 5 s and 30 s was dependent upon the site even though the 5-s and 30-s samples were collected 4 weeks apart.     

Periodontitis lesions are the main source of salivary cytomegalovirus

Background:  Herpesviruses play causal or cooperative roles in childhood infections, tumorigenesis, ulcerogenesis, and periodontitis. Saliva is a common vehicle of herpesvirus horizontal transmission, but the source of salivary herpesviruses remains obscure. To evaluate the significance of periodontal disease in shedding of oral herpesviruses, this study determined the genome-copy counts of human cytomegalovirus (HCMV) and Epstein–Barr virus (EBV) in whole saliva of subjects with periodontitis, gingivitis, or no natural teeth.
Methods:  Whole saliva was collected from 14 periodontitis patients, 15 gingivitis patients and 13 complete denture wearers. The study subjects were systemically healthy and had not received periodontal treatment in the past 3 months. Real-time TaqMan polymerase chain reaction was used to determine the salivary load of HCMV and EBV.
Results:  Salivary HCMV was detected in seven (50%) periodontitis patients, but not in any gingivitis or edentulous subjects ( P  < 0.001). Salivary EBV was detected in 11 (79%) periodontitis patients, in five (33%) gingivitis patients, and in seven (54%) edentulous subjects ( P  = 0.076). Salivary samples showed copy counts of HCMV in the range of 3.3 × 10³–4.2 × 10⁴/ml and of EBV in the range of 3.6 × 10²–1.6 × 10⁹/ml.
Conclusions:  HCMV and EBV are commonly present in the saliva of periodontitis patients. Periodontitis lesions of systemically healthy subjects seem to constitute the main origin of salivary HCMV, but do not comprise the sole source of salivary EBV.     

Herpes viruses in periodontal compromised sites: comparison between HIV-positive and -negative patients

Aim: The objective of this study was to compare the frequency of herpes simplex virus type 1 (HSV‐1), Epstein–Barr virus (EBV) and human cytomegalovirus (HCMV) in subgingival plaque, saliva and peripheral blood of HIV‐positive and‐negative patients with periodontal disease. Material and Methods: Fifty HIV‐positive subjects (23 with gingivitis, 27 with periodontitis) and 50 healthy HIV‐negative patients with chronic periodontitis were included in the study. Parameters of probing depth (PD), clinical attachment level (CAL), gingival index and plaque index were recorded. The samples were processed for viral identification by the nested polymerase chain reaction technique. Results: HCMV was the most prevalent virus in HIV‐positive (82%) and‐negative patients (84%), and the detection in the three samples was similar (p>0.05). HSV‐1 was the least prevalent virus in both groups, being detected in similar frequencies in oral sites and in peripheral blood. EBV‐1 was found more frequently in saliva and subgingival plaque of HIV‐positive patients than in HIV‐negative patients (p0.05). Conclusions: EBV‐1 was more frequently recovered in oral sites of HIV‐positive patients than in HIV‐negative patients.     

Crevicular fluid myeloperoxidase and periodontitis disease

The volume and myeloperoxidase (MPO) activity of gingival crevicular fluid (GCF) collected with filter paper strips for 30 sec from the sulcus of healthy, gingivitis and periodontitis sites of Chinese subjects were measured. MPO/site and MPO/microliter GCF were both greater at gingivitis and periodontitis sites than healthy sites. Enzyme activity was similar at the 2 categories of diseased sites. Mean GCF volume and MPO activity of the samples were calculated for all sites with GI 0, 1, 2 and 0 + 1 irrespective of experimental group assignment. GCF volume MPO/site and MPO/microliter GCF all were greater at GI 2 than GI 0 or 0 + 1. These data indicate that increased GCF MPO previously observed at periodontitis sites is not specific to these sites. Rather, increased GCF MPO likely occurs when additional polymorphonuclear leukocytes enter the sulcus as a result of gingival inflammation. A second sample was obtained from 22 sites 4 weeks after the initial collection. These samples were collected for 5 rather than 30 sec. The GCF volume, MPO/site and MPO/microliter GCF each were greater in samples collected for 30 rather than 5 sec. Correlation coefficients showed that the amount of GCF and MPO activity of the fluid collected for 5 and 30 sec was dependent upon the site even though the samples were collected at different times.