【摘要】：第一部分蘭州地區(qū)口腔門診患者唾液幽門螺桿菌感染情況分析 目的：通過本課題研究,分析蘭州地區(qū)人群唾液幽門螺桿菌(Helicobacter pylori, Hp)感染的細(xì)菌學(xué)資料,了解蘭州地區(qū)人群Hp的感染狀況,為本地區(qū)防治Hp提供科學(xué)依據(jù)。 方法：收集蘭州地區(qū)941例唾液標(biāo)本,行Hp的細(xì)菌學(xué)培養(yǎng),經(jīng)形態(tài)學(xué)、革蘭染色鏡檢、脲酶等生化實驗鑒定Hp的感染狀況。采用卡方檢驗對各分組進(jìn)行檢驗,分析是否存在統(tǒng)計學(xué)意義。 結(jié)果：蘭州地區(qū)人群唾液Hp陽性率為42.72%,不同人群唾液Hp陽性率存在顯著性差異。其中女性為47.89%,顯著高于男性(38.44%)(p=0.004,x2=8.492)；70歲以上年齡組唾液Hp陽性率最高,其次為60歲年齡組,其他各年齡段組間無統(tǒng)計學(xué)差異(p=0.497),而各年齡段口腔唾液Hp生長量無統(tǒng)計學(xué)差異(P=0.086)；按生活環(huán)境分：農(nóng)村人群唾液Hp陽性率(50.93%)顯著高于城市人群(33.99%)(p=0.000,x2=27.551)。 結(jié)論：蘭州地區(qū)唾液Hp陽性率與口腔衛(wèi)生,口腔疾病的發(fā)生,性別,年齡,生活環(huán)境相關(guān)。 第二部分檢測幽門螺桿菌分子生物學(xué)方法的選擇以及口腔幽門螺桿菌的檢測 目的：選擇分子生物學(xué)方法檢測幽門螺桿菌,其較常規(guī)方法更為靈敏快捷。并應(yīng)用于口腔幽門螺桿菌感染的檢測。 方法：采用定性PCR方法和SYBR Green實時熒光定量PCR方法同時檢測甘肅省人民醫(yī)院消化內(nèi)科胃鏡室收集的胃液標(biāo)本,檢測結(jié)果進(jìn)行比較,選擇靈敏性、特異性高的方法應(yīng)用于口腔Hp的檢測。收集甘肅省人民醫(yī)院口腔內(nèi)科138例牙周炎患者牙菌斑標(biāo)本,采用SYBR Green實時熒光定量PCR方法進(jìn)行檢測,分析牙菌斑Hp與牙周炎和消化道疾病之間的相關(guān)性。 結(jié)果：我們建立的SYBR Green實時熒光定量PCR特異性較好,對實驗室其他保存菌種均無擴(kuò)增。對32例胃液標(biāo)本進(jìn)行檢測,我們結(jié)果顯示SYBR Green實時熒光定量PCR方法檢測Hp陽性率為90.63%,而定性PCR方法僅為75.00%。因此,我們使用靈敏性較高的SYBR Green實時熒光定量PCR檢測口腔Hp的存在。運用SYBR Green實時熒光定量PCR檢測138例牙周炎患者牙菌斑標(biāo)本,結(jié)果顯示,牙周健康組牙菌斑Hp陽性率為30.00%,牙周疾病組為74.69%,牙周疾病伴胃病組為92.00%,差異有統(tǒng)計學(xué)意義；且牙菌斑Hp陽性率隨著牙周程病變的程度而增高。 結(jié)論：SYBR Green實時熒光定量PCR可成功檢測胃液和口腔內(nèi)Hp,靈敏度高,特異性強�？谇粌�(nèi)Hp與牙周炎的發(fā)生發(fā)展和消化道疾病密切相關(guān)。
[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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R781;R440

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