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STUDY ON A METHOD FOR MAPPING THE DISTRIBUTION PATTERN OF CARIOGENIC BACTERIA WITHIN DENTAL PLAQUE in vivo USING REAL-TIME PCR

实时荧光定量PCR绘制体内牙菌斑内致龋菌分布模式的方法研究

基本信息

批准号：
14571974
负责人：
KATO Kazuo
金额：
$ 2.5万
依托单位：
AICHI-GAKUIN UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571974/
关键词：
Dental Plaque Cariogenic Bacteria Layered Sample Distribution Real-time PCR polymerase chain reaction

项目摘要

(1)A new method was developed to map the distribution of cariogenic oral streptococci, S.mutans and S.sobrinus, from the outermost to the innermost plaque. The distribution patterns of fractions positive for S.mutans and S. sobrinus varied among the subjects, with a tendency for frequent detection of both species in the outer to middle layers of dental plaque. There were no plaque fractions in which only S.sobrinus was found. This method could be useful to map the distribution of cariogenic microorganisms and to estimate the bacterial ecology for oral biofilm.(2)The antimicrobial effect of SnF_2 gel treatment on dental plaque in vivo was evaluated, using this new method which detects presence of cariogenic streptococci. The results showed that frequencies of layer containing 」 mutans and S.sobrinus were 80% and 37% before, and 94% and 79% after the treatment, respectively, indicating that, contrary to our expectations. the treatment made their habitat in plaque significantly wider.(3)Bacterial density within plaque was analyzed by a quantitative real-time PCR to assess the antimicrobial effect of SnF_2 gel treatment on dental plaque. The density before the treatment peaked close to the outer surface, promptly deceasing towards the interior. However, post treatment levels significantly increased, showing 1010^2 times higher in the outer regions and 10^2-10^3 times higher in the inner regions. Plaque volumes demonstrated a tendency to reduce after the treatment. The results suggested that the application of SnF_2 gel may indeed increase the porosity of plaque, creating a larger habitat for bacteria particularly in deeper regions.

(1)建立了口腔致龋性链球菌、变形链球菌和远缘链球菌从菌斑最外到最内的分布图。变形链球菌和远缘链球菌阳性组分在不同受试者中的分布模式不同，有在牙菌斑的外层和中层频繁检测到这两种细菌的趋势。未发现仅有远缘链霉菌的菌斑组分。利用这种检测致龋性链球菌的新方法，在体内评价了SNF2凝胶处理对牙菌斑的抑菌效果。结果表明，治疗前蛋鸡中含有变种和远缘链霉菌的频率分别为80%和37%，治疗后分别为94%和79%，这与我们的预期相反。采用实时定量聚合酶链式反应分析菌斑内细菌密度，评价三氟化钠凝胶处理对牙菌斑的抑菌效果。处理前的密度在接近外表面时达到峰值，向内迅速下降。然而，治疗后的水平显著增加，表现为外部区域高出10102倍，内部区域高出10^2-10^3倍。治疗后斑块体积有减少的趋势。结果表明，SNF2凝胶的应用确实可以增加菌斑的孔隙度，为细菌创造更大的栖息地，特别是在更深的区域。