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  本文選題：實時熒光定量PCR + 慢性牙周炎��； 參考：《蘭州大學》2014年碩士論文

【摘要】：牙周病是導致成年人牙齒缺失的首要原因,在國人口腔中普遍存在的一種疾病,卻不為重視。牙周病病原菌以厭氧菌為主,例如,牙齦卟啉單胞菌(Porphyromonas gingivalis, Pg)、伴放線放線桿菌(Actinobacillus actinomycetem-comomitans, Aa)、福賽坦氏菌(Tannerella forsythia,Tf)等,快速檢測病灶組織病原菌含量對疾病的診斷,治療效果的評估意義非凡。培養(yǎng)法是一種古老且常被采用的微生物檢測手段,但是口腔厭氧菌在培養(yǎng)基中極其難以生長,況且目前大多數(shù)的分子生物學研究手段只是針對口腔病原菌的定性研究,如常規(guī)PCR,然而熒光定量PCR可上升到樣本中目標菌株量的探究,并且具有靈敏、特異、快速等優(yōu)勢。唾液是人體一種重要的體液,組成成分復雜,容易采集,對患者無侵入性,因此,唾液診斷在牙周病、齲病的早期發(fā)現(xiàn)、判定療效、及預后評估方面具有很好的應用前景。 目的：本實驗擬用SYBRgreen模式的實時熒光定量PCR技術(shù),針對Pg特異基因Arg-gingipain,對牙周炎患者,牙周健康人群唾液中Pg進行定量檢測,比較在各組人群中Pg數(shù)量的差異,探討Pg在牙周炎患者中的分布及其致病機理,從而為該病診斷治療和療效評估提供依據(jù)。 方法：利用SYBRgreen模式的實時定量PCR方法檢測了20例慢性牙周炎和20例牙周健康者唾液內(nèi)Pg,經(jīng)t檢驗分析比較在各組人群中Pg定植的差異。 結(jié)果：慢性牙周炎患者唾液中,Pg的檢出數(shù)目對數(shù)值1.01±0.77,檢出率為85%,其中,女性患者Pg的檢出數(shù)目對數(shù)值1.02±0.83,男性患者Pg的檢出數(shù)目對數(shù)值1.00±0.76；牙周健康者唾液中,Pg的檢出數(shù)目對數(shù)值0.35±0.11,檢出率為10%,女性患者Pg的檢出數(shù)目對數(shù)值0.34±0.10,男性患者Pg檢出數(shù)目對數(shù)值0.36±0.12。Pg在慢性牙周炎患者唾液中的檢出量明顯高于牙周健康組(P0.05),牙周炎男女之間唾液中Pg的檢出數(shù)目無統(tǒng)計學差異(P0.05)。 結(jié)論：唾液診斷是一種可靠的微生物檢出方式,值得推廣并使之應用于臨床疾病的輔助診斷。驗證了牙齦卟啉單胞菌與慢性牙周病發(fā)生發(fā)展的相關(guān)性,同時發(fā)現(xiàn)Pg的檢出量與性別沒有明顯相關(guān)性。
[Abstract]:Periodontal disease is the leading cause of tooth loss in adults. The main pathogenic bacteria of periodontal disease were anaerobic bacteria, such as Porphyromonas gingivalis, PgGV, Actinobacillus actinomycetem-comomitans, Aaer, Tannerella forsythiaTf. etc. Culture is an ancient and often used method for microbial detection, but oral anaerobes are extremely difficult to grow in the medium, and most of the current molecular biological research methods are only qualitative studies of oral pathogens. As with conventional PCR, however, fluorescence quantitative PCR can increase to the target strain quantity in the sample, and has the advantages of sensitivity, specificity, rapidity and so on. Saliva is an important humoral fluid of human body, which is complex in composition, easy to collect, and not invasive to patients. Therefore, saliva diagnosis has a good application prospect in the early detection of periodontal disease, caries, evaluation of curative effect and prognosis evaluation. Objective: the aim of this study was to quantitatively detect PG in saliva of periodontitis patients and healthy people by real-time fluorescence quantitative PCR (PCR) based on SYBRgreen model. To investigate the distribution of PG in periodontitis and its pathogenic mechanism, and to provide basis for diagnosis, treatment and evaluation of curative effect of periodontitis. Methods: SYBRgreen real-time quantitative PCR method was used to detect the salivary Pg in 20 cases of chronic periodontitis and 20 cases of periodontal healthy subjects, and the difference of PG colonization in each group was compared by t test. Results: the logarithmic value of Pg in saliva of patients with chronic periodontitis was 1.01 鹵0.77, and the detection rate was 85. The logarithmic value of detection of PG in female patients was 1.02 鹵0.83and that in male patients was 1.00 鹵0.76.The logarithmic value of detection of PG in saliva of periodontal health patients was 0.35 鹵0.11, and the detection rate was 10. The logarithmic value of detection of PG in female patients was 0.34 鹵0.10, and that in male patients was 0.34 鹵0.10. The detectable amount of logarithmic value of 0. 36 鹵0.12.Pg in saliva of chronic periodontitis patients was significantly higher than that of healthy periodontitis group (P0. 05). There was no significant difference in the detection number of PG in saliva between male and female patients with periodontitis (P 0. 05). Conclusion: saliva diagnosis is a reliable method for microbial detection, which is worth popularizing and applying to the auxiliary diagnosis of clinical diseases. The correlation between Porphyromonas gingivalis and the occurrence and development of chronic periodontal disease was verified.
【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R781.42

【參考文獻】

相關(guān)期刊論文 前10條

1 袁亞男;劉文忠;;實時熒光定量PCR技術(shù)的類型、特點與應用[J];中國畜牧獸醫(yī);2008年03期

2 王愛民;;實時熒光定量PCR(TaqMan)法測定外源基因的拷貝數(shù)[J];廣西植物;2009年03期

3 潘春玲,潘亞萍,林莉,趙戩,張冬梅;牙齦卟啉單胞菌在齦下菌斑和頰黏膜中的檢測[J];華西口腔醫(yī)學雜志;2005年05期

4 王麗;楊禾;吳亞菲;;紙尖法和刮匙法對齦下牙周致病菌檢出比較研究[J];臨床口腔醫(yī)學雜志;2008年12期

5 余琦;李寧;李彩云;宋業(yè)穎;楊江輝;劉成龍;羅煥超;;應用熒光定量PCR法檢測病理組織中幽門螺旋桿菌感染[J];局解手術(shù)學雜志;2012年05期

6 劉小榮;張笠;王勇平;;實時熒光定量PCR技術(shù)的理論研究及其醫(yī)學應用[J];中國組織工程研究與臨床康復;2010年02期

7 平飛云;孫鋼;何虹;;SELDI-TOF-MS技術(shù)在口腔黏膜癌前病變和口腔鱗癌的唾液診斷模型中的應用[J];中國病理生理雜志;2010年01期

8 孫德剛;馬晟利;楊俊玉;孫雪梅;吳雙燕;;健康青少年口腔優(yōu)勢菌對牙齦卟啉單胞菌拮抗作用的實驗研究[J];中國微生態(tài)學雜志;2006年04期

9 金笑一,楊圣輝,張春梅,楊新海,王申五;RPC技術(shù)檢測兒童齦炎中的卟啉單胞菌[J];中國微生態(tài)學雜志;1999年02期

10 任小紅;;唾液的分泌及影響因素[J];中國實用醫(yī)藥;2009年20期

，

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