牙周炎及胃病患者牙菌斑中的幽门螺杆菌

目的 明确牙周炎及胃病患者的牙菌斑中是否存在幽门螺杆菌（Ｈｐ）及Ｈｐ是否为细胞毒素相关基因Ａ阳性。方法 利用尿素酶Ｃ基因和ｃａｇＡ设计引物,通过聚合酶链反应（ＰＣＲ）检测口腔中的Ｈｐ。选择１３例胃病及１０例牙周炎患者,每例患者选６个有牙龈炎症的牙位取龈上、龈下菌斑,共计２７６份样本用于ＰＣＲ检测。结果 胃病组１１例患者和牙周炎组全部病例均少有１份菌斑样本检出Ｈｐ,其中尿素酶Ｃ基因在胃病组的阳性率为     

不同方法检测牙菌斑中幽门螺杆菌的比较

目的 探讨用二氧化硅微粒法提取DNA检测口腔标本中幽门螺杆菌可行性及临床应用的意义。方法 选择50例因上腹部症状且有不同程度牙周炎的胃镜检查者50例。每例均取口腔标本。比较二氧化硅微粒法与DNA快提法两种不同方法提取胃及口腔样本中的DNA，引物选用尿素酶C基因和CagA基因进行PCR扩增。结果 SiO2微粒法提取DNA检测菌斑和含漱液中的幽门螺杆菌与Instagene Matrix提取DNA(Bio-rad)比较灵敏度为93．8％(30／32)，特异性为100％(68／68)。结论 SiO2微粒法提取菌斑和含漱液中的DNA操作简便，可作为Instagene Matrix替代方法检测口腔中的Hp。     

高压氧对口腔牙菌斑幽门螺杆菌的作用

目的：寻找一种较为理想的根除口腔幽门螺杆菌（Hp)的方法。方法：将120例口腔牙菌斑Hp-PCR检测阳性的Ⅱ度牙周炎患者分为A、B、C、D4组,治疗1、3、6月后分别对所有患者牙周参数和龈下菌斑进行Hp-PCR检测并比较检测结果。结果：4组Hp阳性率分别为96.7%、80.0%、73.3%和0,有显著性差异（P＜0.05),其中以D组最低（P＜0.05)。结论：高压氧对口腔Hp有明显抑制或杀灭作用,高压氧配合治疗,有助于口腔Hp的根除和与Hp有关的疾病的治疗。     

幽门螺杆菌在牙髓中的分布研究

目的：检测人牙髓标本中Hp的存在情况，探计Hp与牙髓病变的相关性以及Hp的传播途径和传播方式。方法：本实验分为5组，共计79例标本，前4组取根髓，第5组取龈下牙石。采用聚合酶链反应(PCR)和Hp快速诊断试剂盒两种方法检测不同病情下人牙髓和牙石标本中的Hp。结果：经x^2检验各牙髓组间Hp检出率无明显性差异；而牙髓组与龈下牙石组间Hp检出率均有显著性差异。结论：发现在人牙髓中存在Hp，牙髓中幽门螺杆菌的存在可能是Hp不易清除和相关疾病易于复发的原因。     

Pyrosequencing 检测口腔与胃中的幽门螺杆菌16S rDNA V1区基因序列

目的 通过Pyrosequencing检测口腔与胃幽门螺杆菌16SrDNA V1区序列基因片段分析，以进一步验证口-口传播的的假设。方法 选择18例患有慢性中度牙周炎且胃镜活检Hp感染阳性患者及其中10例患者的家属。提取患者胃、牙菌斑和含漱液共74例样本中的DNA，PCR扩增阳性后pyrosequecing检测16SrDNA V1区序列基因片段。结果患者胃和口腔Hp及其家属口腔Hp的序列相比较，有0～1个碱基不同。结论 口腔中的Hp与胃内Hp16SrDNA V1区基因型比较95．8％～100%的同源性，口腔可能为Hp的聚集地。口腔Hp可能与胃Hp感染的复发或再感染有关。     

口腔中的幽门螺杆菌

幽门螺杆菌（Ｈｐ）是革兰氏阴性微需氧菌，具多形性，通常存在于胃肠道上部。应用培养、生化分析和核酸技术在牙菌斑和唾液中分离或检测到了Ｈｐ。Ｈｐ的传播途径尚不清楚，以口－口传播为主。口腔中的Ｈｐ可能是胃部Ｈｐ再感染的重要储存库，也可能是引起口臭的病原菌之一，并可能与某些口腔疾病如复发性口疮的发生有关。     

牙菌斑中幽门螺杆菌与胃病的关系

目的 :分离培养口腔牙菌斑中的幽门螺杆菌 (Helicobackerpylori,Hp)并探讨它与Hp相关性慢性胃病之间的相互关系。方法 :采用细菌培养的方法检测 40例体检者的龈下牙菌斑和胃黏膜中的Hp。 结果 :10例 ( 2 7.0 3 % )慢性胃病患者的胃黏膜Hp培养阳性 ,其中 5例 ( 13 .5 1% )牙菌斑Hp培养阳性。 3例胃黏膜Hp培养阴性的非慢性胃病患者中有 1例牙菌斑Hp培养阳性。 结论 :牙菌斑是Hp的另一个重要储存库 ,口腔Hp的存在可能与慢性胃病有一定的相关性。     

口腔幽门螺杆菌对胃幽门螺杆菌根除率的影响

目的　明确口腔幽门螺杆菌 (Helicobacterpylori,Hp)对胃幽门螺杆菌根除率的影响。方法　选择 10 2例有上腹部症状、并经全口牙周检查有不同程度牙周炎的患者进行胃镜检查 ,每例均取口腔标本进行Hp检测。用二氧化硅微粒法提取所有样本中的DNA ,引物选用尿素酶C基因和CagA基因进行PCR扩增。结果　胃Hp阳性组的口腔Hp检出率 (n =5 8,4 3.1% )高于胃Hp阴性组(n =4 4 ,2 2 .7% ) ,两者比较差异有显著性 (P <0 .0 5 )。 5 8例胃Hp阳性患者经药物治疗后 4周进行胃Hp的复查 ,其中口腔Hp阳性者的胃Hp根除率 (6 4 0 % ,16 / 2 5 )低于口腔Hp阴性者 (72 .7% ,2 4 / 33) ;治疗后 1年 ,口腔Hp阳性者的胃Hp根除率 (36 .0 % )与Hp阴性者的根除率 (6 3.6 % )相比差异有显著性 (P <0 .0 5 )。结论　胃Hp的根除率受口腔Hp存在的影响 ;口腔Hp阳性者的胃Hp根除率显著低于口腔Hp阴性者根除率     

用PCR技术同时检测牙菌斑、唾液及胃粘膜中的幽门螺杆菌

目的：通过同时测定牙菌斑、唾液及胃粘膜中幽门螺杆菌（Hp）以了解口腔中Hp的状态及其与胃粘膜病变的关系。方法：采用PCR技术同时测定牙菌斑、唾液及胃粘膜中的幽门螺杆菌。结果：47例胃粘膜Hp阳性的胃窦炎患者中17例（36．2％）牙菌斑中检出Hp;31例（65．9％）唾液标本Hp阳性。而13例胃粘膜Hp阴性者中仍有2例牙菌斑中检出的Hp,1例呼液中检出Hp。胃粘膜Hp阳性和阴性者之间牙菌斑及唾液Hp检出率的差异有显著性（P＜0．01）。结论：口腔中的Hp可能与胃粘膜中的Hp存在着一定的病因学联系。     

口腔健康教育对牙周参数和幽门螺杆菌在牙菌斑中分布的影响

目的 探讨口腔卫生教育对牙周参数和幽门螺杆菌（Hp）在牙菌斑中分布的影响。方法 自愿受检人群进行口腔健康教育，统一指导漱口、刷牙并口腔洁治，抽样观察牙周参数和菌斑Hp阳性率的变化。结果 口腔健康教育后，牙周参数和Hp阳性率均有显著性降低（P＜0.01），其效果以洁治教育后最为显著（P＜0.001），但正确的刷牙是保持口腔卫生最基本的方法。结论 进行口腔健康教育不仅可以减少口腔疾病，对减小胃病的发病与复发亦有重要意义。     

Prevalence of Helicobacter pylori detected by polymerase chain reaction in the oral cavity of periodontitis patients

Helicobacter pylori is an important gastrointestinal pathogen associated with gastritis, peptic ulcers, and an increased risk of gastric carcinoma. The oral cavity has been indicated as a possible H. pylori reservoir, and may therefore be involved in the reinfection of the stomach which sometimes follows treatment of H. pylori infection. The objective of the present study was to evaluate the prevalence of H. pylori as detected by polymerase chain reaction (PCR) in the oral cavity of periodontitis patients testing positive for this bacterium in the stomach. Thirty adult patients with alterations of the superior digestive tract, testing urease positive after endoscopy and biopsy, were selected. A full-mouth periodontal examination was performed in every patient and the subjects were allocated to two groups: gingivitis (15 patients) and chronic periodontitis (15 patients). Plaque and saliva samples collected from each patient were stored in 0.5 ml of TE buffer. DNA was extracted from the samples by the boiling method and was evaluated for the presence of H. pylori using the PCR method. JW 22/23 primers were used. The DNA of ATCC H. pylori 43629 (positive control) and water (negative control) were used for controlling the reactions. Of the 30 evaluated patients, 13 (43.3%) harbored H. pylori in the mouth. The bacterium was not found on the dorsum of the tongue of any patient, but was found in saliva in three patients (10%), in the supragingival plaque in six patients (20%), and in the subgingival plaque in eight patients (26.6%). The presence of H. pylori was similar in the gingivitis and chronic periodontitis groups. In conclusion, a high percentage of patients harbored H. pylori in their mouth. The bacterium was detected in saliva, supragingival and subgingival plaque, suggesting that these sites may be considered reservoirs for H. pylori in urease-positive patients.     

慢性牙周炎与口腔幽门螺杆菌的相关性研究

目的：探讨慢性牙周炎患者与其口腔中的幽门螺杆菌（helicobacter pylori,HP）的相关性。方法：依据HP特异的尿素酶C和cagA基因设计引物,建立PCR方法。检测37例牙周炎患者口腔菌斑和含漱液中的HP,同时设立牙周健康对照组,比较两组的检测结果。结果：尿素酶C基因在牙周炎组的阳性率为64.8%,健康组为38.5%;尿素酶C基因和cagA基因共同阳性率在牙周炎组为32.4%,健康组为7.7%;两组差异有统计学意义;且轻型牙周炎患者口腔HP阳性率低于中、重型患者。龈下菌班HP阳性率高于龈上菌斑。结论：HP参与了慢性牙周炎的致病过程,抑或是加强了其它因素的致病过程,HP与牙周炎有相关性。     

慢性胃病患者口腔与胃内幽门螺杆菌研究

目的：研究慢性胃病患者口腔中是否存在幽门螺杆菌（Hp）,并分析胃病患者牙周状况与幽门螺杆菌的关系。方法：依据特异的尿素酶C基因和cagA基因设计引物,建立聚合酶链反应（PCR）方法。检测65例慢性胃病患者不同牙位龈上和龈下菌斑中的Hp。结果：65例慢性胃病患者胃黏膜标本中,PCR检测阳性标本58例（89．3％）,龈下菌斑Hp阳性率（47．7％）,高于龈上菌斑中的Hp阳性率（26．2％）。牙周袋深度（PD）≥4mm部位菌斑中Hp检出率显著高于PD〈4mm的部位（P〈0．05）。结论：口腔Hp是慢性胃病患者Hp感染的重要来源。     

Absence of Helicobacter pylori in subgingival samples determined by polymerase chain reaction

The polymerase chain reaction was used for the detection of Helicobacter pylori from subgingival plaque in 336 periodontitis patients. A pair of primers derived from the H. pylori urease gene A served to amplify a targeted 411-bp fragment of genomic DNA. This technique permitted the detection of as few as 60 H. pylori cells. Paper point samples from 3 deep periodontal pockets per patient were immersed in 1 ml of phosphate-buffered saline or distilled water, DNA was solubilized by detergent/protease method, 3.7 μl or 37 μl of lysate supernatant was used as template, and the amplification product was analyzed in 1% agarose gel containing ethidium bromide. Each experiment included purified DNA and cell lysate of H. pylori as positive controls. The presence of bacteria in the sample was verified by a primer pair common to prokaryote 16S rRNA. The present study did not reveal the specific polymerase chain reaction amplification product characteristic of H. pylori. We conclude that periodontal pockets do not constitute a natural reservoir for H. pylori.     

Detection of putative periodontal pathogens in non-insulin-dependent diabetes mellitus and non-diabetes mellitus by polymerase chain reaction

It has been assumed that there is a relationship between periodontal diseases and diabetes mellitus, however the putative periodontal microorganisms in non-diabetes mellitus (non-DM) individuals and non-insulin-dependent diabetes mellitus (NIDDM) patients have not been well studied. In this study, the detection rates of 5 putative periodontal pathogens: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Eikenella corrodens, Treponema denticola, and Candida albicans by polymerase chain reaction (PCR) between NIDDM and non-DM adults were compared. A total of 246 adults were randomly recruited and periodontal parameters including: plaque index (P1I), gingival index (GI), probing depth (PD) and attachment level (AL) were recorded. Subgingival plaque samples were collected by sterile curettes from the most diseased and healthy sites based on PD and AL. The differences in periodontal parameters and microbiological data in healthy and diseased sites between non-DM and NIDDM patients were compared by chi-square analysis. The results showed no significant differences in age, gender, GI, P1I, PD, and prevalence of the 5 microorganisms between the NIDDM and the non-diabetic groups. However, except for A. actinomycetemcomitans, the prevalence of the periodontal microorganisms tested was significantly higher (p <0.001) in diseased sites than in the healthy sites in both groups. The P1I, GI, PD and AL were significantly higher in T. denticola positive sites than in negative sites. The results suggested that P. gingivalis, T. denticola, E. corrodens and C. albicans may play important roles in the periodontitis of both NIDDM and non-DM individuals, however the etiology of periodontitis in both groups may not be different from each other.     

Detection of Helicobacter pylori in various oral lesions by nested polymerase chain reaction (PCR)

Mravak-Stipetic M, Gall-Troselj K, Lukac J, Kusic Z, Pavelic K, Pavelic J: Detection of Helicobacter pylori in various oral lesions by nested polymerase chain reaction (PCR). J Oral Pathol Med 1998; 27: 1–3. © Munksgaard, 1998.
Nested PCR was used for the detection of Helicobacter pylori DNA in specimens collected from seven different topographic sites in the oral cavity. Out of 161 patients, only 21 (13.04%) were positive. There was no correlation between H. pylori status and patient diagnosis and age. No preferential site for bacterial colonization was found in the oral cavity, nor was an association established between a bacterial presence and ulcerated versus non-ulcerated lesions. The results indicate that the oral mucosa does not appear to represent a preferred site of colonization for H. pylori. Furthermore, the evidence presented in this paper suggests that H. pylori is not pathogenic in the oral cavity, nor is it associated with common oral pathologic processes.     

幽门螺杆菌在使用的牙刷上分布的研究

目的：为探讨幽门螺杆菌（helicobacter pylori,Hp)的感染方式和感染途径。本实验对不同牙周情况和胃病患病情况的自愿者使用的牙刷上Hp的分布进行研究。方法 以牙刷毛基部沉积物为标本,采用聚合酶链反应（polymerase chain reaction,PCR)方法提取Hp染色体NDA,对其特异性片段进行诊断。结果：牙周状况较好者,无胃病者使用的牙刷Hp 检出率较低;并且牙刷使用时间越长,Hp在牙刷毛基部沉积物上孳生的可能越大（P＜0．01）,同时牙刷的正确使用与保护对Hp检出率亦有非常显著的影响（P＜0．05）,结论：牙刷上Hp的分布说明口腔可能是Hp传播的一条通道,因此牙刷使用时间不宜过长,另外养成良好的口腔卫生习惯也十分重要。     

Absence of Helicobacter pylori in the oral cavity of 10 non-dyspeptic subjects demonstrated by real-time polymerase chain reaction

Helicobacter pylori plays a significant role in gastric disease. However, the presence of this bacterium in the oral cavity remains controversial. The aim of the present study was to detect and quantify H. pylori in 29 different sites of the oral cavity in non-dyspeptic subjects by means of real-time polymerase chain reactions (PCR). Ten subjects without gastric symptoms were studied. Samples from unstimulated saliva, three sites of the tongue, oral mucosa, and 12 sites of both supragingival and subgingival plaque were collected from each subject. DNA was extracted from the oral samples and analysed for the presence of H. pylori by real-time PCR (LightCycler) using JW23/22 primers which targeted the 16S rRNA gene. DNA from H. pylori DSM 4867 was used as a positive control. Amplification efficiency for the LightCycler 2.0 runs ranged from 1.8 to 2.4. Melting curve analysis identified all the positive control capillaries, which contained H. pylori reference DNA, as a single and narrow peak at a melting temperature between 84.5 and 84.9 degrees C. All the negative control capillaries with no template control and the 29 oral samples from each subject showed either no melting peaks or broad melting peaks below 80 degrees C, which were considered as primer dimers. Therefore, H. pylori was not detected from any of the 290 oral samples. As a conclusion, H. pylori seems not to be permanently present in the oral cavity of a non-dyspeptic population.     

Comparative analysis of putative periodontopathic bacteria by multiplex polymerase chain reaction

Background and Objective:  The polymerase chain reaction (PCR) has been applied for the rapid and specific detection of periodontopathic bacteria in subgingival plaque and is potentially of clinical benefit in the diagnosis and treatment of periodontitis subjects. However, several technical points need to be modified before the conventional PCR detection system can be used by clinicians.
Material and Methods:  To develop a PCR-based technique more applicable for clinical use than conventional PCR, we established a multiplex PCR for five putative periodontopathic ( Treponema denticola , Porphyromonas gingivalis , Aggregatibacter actinomycetemcomitans , Prevotella intermedia and Tannerella forsythia ) and two nonperiodontopathic ( Streptococcus sanguinis and Streptococcus salivarius ) species of bacteria using whole-plaque suspension as templates, and detected bacteria in subgingival plaque taken from 85 subjects at the supportive periodontal therapy stage after active periodontal treatments.
Results:  Among putative periodontopathic bacteria, the detection frequency of T. denticola and P. gingivalis was elevated in parallel with higher probing pocket depth and clinical attachment loss, and had 4.2–14.1 times increasing odds of the clinical parameters tested. Detection of any of the five species of putative periodontopathic bacteria markedly increased the odds ratio of a higher probing pocket depth, clinical attachment loss and bleeding on probing.
Conclusion:  The multiplex PCR system developed in this study enabled the detection of all the bacteria under investigation in one reaction tube in a less time- and labor-intensive manner than conventional PCR. These results support the potential clinical use of multiplex PCR for detecting periodontopathic bacteria and for evaluating therapeutic strategies and predicting the prognosis for each subject.