口腔唾液牙龈卟啉单胞菌的快速微量检测

发布时间：2018-04-29 06:21

本文选题：实时荧光定量PCR + 慢性牙周炎　；参考：《兰州大学》2014年硕士论文

【摘要】：牙周病是导致成年人牙齿缺失的首要原因,在国人口腔中普遍存在的一种疾病,却不为重视。牙周病病原菌以厌氧菌为主,例如,牙龈卟啉单胞菌(Porphyromonas gingivalis, Pg)、伴放线放线杆菌(Actinobacillus actinomycetem-comomitans, Aa)、福赛坦氏菌(Tannerella forsythia,Tf)等,快速检测病灶组织病原菌含量对疾病的诊断,治疗效果的评估意义非凡。培养法是一种古老且常被采用的微生物检测手段,但是口腔厌氧菌在培养基中极其难以生长,况且目前大多数的分子生物学研究手段只是针对口腔病原菌的定性研究,如常规PCR,然而荧光定量PCR可上升到样本中目标菌株量的探究,并且具有灵敏、特异、快速等优势。唾液是人体一种重要的体液,组成成分复杂,容易采集,对患者无侵入性,因此,唾液诊断在牙周病、龋病的早期发现、判定疗效、及预后评估方面具有很好的应用前景。目的：本实验拟用SYBRgreen模式的实时荧光定量PCR技术,针对Pg特异基因Arg-gingipain,对牙周炎患者,牙周健康人群唾液中Pg进行定量检测,比较在各组人群中Pg数量的差异,探讨Pg在牙周炎患者中的分布及其致病机理,从而为该病诊断治疗和疗效评估提供依据。方法：利用SYBRgreen模式的实时定量PCR方法检测了20例慢性牙周炎和20例牙周健康者唾液内Pg,经t检验分析比较在各组人群中Pg定植的差异。结果：慢性牙周炎患者唾液中,Pg的检出数目对数值1.01±0.77,检出率为85%,其中,女性患者Pg的检出数目对数值1.02±0.83,男性患者Pg的检出数目对数值1.00±0.76；牙周健康者唾液中,Pg的检出数目对数值0.35±0.11,检出率为10%,女性患者Pg的检出数目对数值0.34±0.10,男性患者Pg检出数目对数值0.36±0.12。Pg在慢性牙周炎患者唾液中的检出量明显高于牙周健康组(P0.05),牙周炎男女之间唾液中Pg的检出数目无统计学差异(P0.05)。结论：唾液诊断是一种可靠的微生物检出方式,值得推广并使之应用于临床疾病的辅助诊断。验证了牙龈卟啉单胞菌与慢性牙周病发生发展的相关性,同时发现Pg的检出量与性别没有明显相关性。
[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 卤0.77, and the detection rate was 85. The logarithmic value of detection of PG in female patients was 1.02 卤0.83and that in male patients was 1.00 卤0.76.The logarithmic value of detection of PG in saliva of periodontal health patients was 0.35 卤0.11, and the detection rate was 10. The logarithmic value of detection of PG in female patients was 0.34 卤0.10, and that in male patients was 0.34 卤0.10. The detectable amount of logarithmic value of 0. 36 卤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.
【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】：R781.42

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