口腔幽门螺杆菌的感染状况分析及检测方法比较

发布时间：2018-12-23 12:54

【摘要】：第一部分兰州地区口腔门诊患者唾液幽门螺杆菌感染情况分析目的：通过本课题研究,分析兰州地区人群唾液幽门螺杆菌(Helicobacter pylori, Hp)感染的细菌学资料,了解兰州地区人群Hp的感染状况,为本地区防治Hp提供科学依据。方法：收集兰州地区941例唾液标本,行Hp的细菌学培养,经形态学、革兰染色镜检、脲酶等生化实验鉴定Hp的感染状况。采用卡方检验对各分组进行检验,分析是否存在统计学意义。结果：兰州地区人群唾液Hp阳性率为42.72%,不同人群唾液Hp阳性率存在显著性差异。其中女性为47.89%,显著高于男性(38.44%)(p=0.004,x2=8.492)；70岁以上年龄组唾液Hp阳性率最高,其次为60岁年龄组,其他各年龄段组间无统计学差异(p=0.497),而各年龄段口腔唾液Hp生长量无统计学差异(P=0.086)；按生活环境分：农村人群唾液Hp阳性率(50.93%)显著高于城市人群(33.99%)(p=0.000,x2=27.551)。结论：兰州地区唾液Hp阳性率与口腔卫生,口腔疾病的发生,性别,年龄,生活环境相关。第二部分检测幽门螺杆菌分子生物学方法的选择以及口腔幽门螺杆菌的检测目的：选择分子生物学方法检测幽门螺杆菌,其较常规方法更为灵敏快捷。并应用于口腔幽门螺杆菌感染的检测。方法：采用定性PCR方法和SYBR Green实时荧光定量PCR方法同时检测甘肃省人民医院消化内科胃镜室收集的胃液标本,检测结果进行比较,选择灵敏性、特异性高的方法应用于口腔Hp的检测。收集甘肃省人民医院口腔内科138例牙周炎患者牙菌斑标本,采用SYBR Green实时荧光定量PCR方法进行检测,分析牙菌斑Hp与牙周炎和消化道疾病之间的相关性。结果：我们建立的SYBR Green实时荧光定量PCR特异性较好,对实验室其他保存菌种均无扩增。对32例胃液标本进行检测,我们结果显示SYBR Green实时荧光定量PCR方法检测Hp阳性率为90.63%,而定性PCR方法仅为75.00%。因此,我们使用灵敏性较高的SYBR Green实时荧光定量PCR检测口腔Hp的存在。运用SYBR Green实时荧光定量PCR检测138例牙周炎患者牙菌斑标本,结果显示,牙周健康组牙菌斑Hp阳性率为30.00%,牙周疾病组为74.69%,牙周疾病伴胃病组为92.00%,差异有统计学意义；且牙菌斑Hp阳性率随着牙周程病变的程度而增高。结论：SYBR Green实时荧光定量PCR可成功检测胃液和口腔内Hp,灵敏度高,特异性强。口腔内Hp与牙周炎的发生发展和消化道疾病密切相关。
[Abstract]:Part I Analysis of Helicobacter pylori infection in saliva of outpatients in Lanzhou area objective: to analyze the bacteriological data of Helicobacter pylori (Helicobacter pylori, Hp) infection in saliva of population in Lanzhou area. To understand the infection status of Hp in Lanzhou area and to provide scientific basis for prevention and treatment of Hp in this area. Methods: 941 saliva samples from Lanzhou area were collected. The bacterial culture of Hp was performed. The infection status of Hp was identified by morphology, Gram staining, urease and other biochemical tests. Chi-square test was used to test each group to analyze whether there was statistical significance. Results: the positive rate of salivary Hp in Lanzhou area was 42.72, and the positive rate of salivary Hp was significantly different among different populations. The percentage of female was 47.89, which was significantly higher than that of male (38.44%) (p0.004% x 2.492). The positive rate of Hp in saliva was the highest in the group over 70 years old, followed by the age group of 60 years old. There was no statistical difference among other age groups (p0. 497), but there was no significant difference in the growth of Hp in oral saliva of different age groups (P0. 086). According to living environment, the positive rate of saliva Hp in rural population (50.93%) was significantly higher than that in urban population (33.99%). Conclusion: the positive rate of saliva Hp in Lanzhou area is related to oral health, oral disease, sex, age and living environment. The second part is the selection of molecular biological methods for detection of Helicobacter pylori and the detection of Helicobacter pylori in oral cavity. And applied to the detection of Helicobacter pylori infection in oral cavity. Methods: using qualitative PCR method and SYBR Green real-time fluorescence quantitative PCR method to detect gastric juice samples collected from gastroscope of Department of Gastroscopy, Gansu Provincial people's Hospital, and to compare the results and select sensitivity. The method with high specificity was applied to the detection of oral Hp. The plaque samples of 138 patients with periodontitis were collected from Department of Stomatology, people's Hospital of Gansu Province. The correlation between Hp and periodontitis and digestive tract diseases was analyzed by SYBR Green real-time fluorescence quantitative PCR. Results: the SYBR Green real-time quantitative PCR has good specificity and has no amplification to other preserved strains in laboratory. The positive rate of Hp detected by SYBR Green real-time fluorescence quantitative PCR method was 90.63, while that by qualitative PCR method was 75.00%. Therefore, we use SYBR Green real-time quantitative PCR with high sensitivity to detect the presence of oral Hp. The positive rates of Hp in periodontal plaque, periodontal disease and gastropathy were 30.00,74.69 and 92.00 respectively in healthy periodontal group, periodontal disease group and gastric disease group, respectively, using SYBR Green real-time fluorescence quantitative PCR. The results showed that the positive rate of Hp in periodontal healthy group was 30.00,74.69 in periodontal disease group, and 92.00 in periodontal disease with stomach disease group. The difference was statistically significant. The positive rate of Hp in dental plaque increased with the degree of periodontal disease. Conclusion: SYBR Green real-time fluorescence quantitative PCR can detect Hp, in gastric juice and oral cavity with high sensitivity and specificity. Oral Hp is closely related to periodontitis and digestive tract diseases.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781;R440

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