Molecular Analysis of Infection in Advanced Caries

描述：龋损向牙本质基质的延伸破坏了牙齿的生理和功能的完整性，并对牙髓组织造成了越来越严重的伤害。与这一过程相关的细菌还没有被很好地了解。目前，没有可靠的手段来确定龋损下牙髓组织的状态，也没有改进的治疗方法可用。这严重损害了目前对这种临床重要疾病的诊断和治疗方案的有效性。 初步研究使用了基于16S rDNA的通用细菌扩增子来确定龋齿牙本质的主要细菌种群。这项工作还得到了实时聚合酶链式反应定量细菌DNA的补充，以确定通过系统发育分析鉴定的细菌种类的水平。一组以厌氧菌为主的复杂阵列被证明是具有特征的乳杆菌和普雷沃特氏菌。占主导地位。通过开发丰富低丰度感兴趣物种种群分析的方法，将加强对正在发育的病变的微生物成分的进一步剖析。获得的序列信息将被用来设计特定的寡核苷酸探针，通过荧光原位杂交结合共聚焦显微镜来确定病变内生物的空间定位。 实验分析将利用提取和原位两种方法提供高分辨率的牙髓细菌入侵机制的信息，并将其与牙髓的组织病理学反应联系起来。本研究获得的数据将为了解龋病过程的扩展提供客观依据，对预测牙髓反应，包括牙髓死亡和牙髓感染具有潜在的诊断价值。

DESCRIPTION: Extension of the carious lesion into the matrix of dentine undermines the physical and functional integrity of the tooth and provides an increasingly noxious insult to the underlying pulp tissue. The bacteria associated with this process are not well understood. Currently, there are no reliable means of identifying the status of the pulp tissue underlying a carious lesion and no enhanced therapeutic modalities are available. This seriously impairs the efficacy of current diagnostic and treatment regimes for this clinically important disease. Preliminary studies have used a universal bacterial amplicon based on 16S rDNA to define the major bacterial populations of carious dentine. This work was complemented by real-time polymerase chain reaction quantification of bacterial DMA to determine levels of bacterial species identified by phylogenetic analysis. A complex array of predominantly anaerobic bacteria was demonstrated with characteristic Lactobacillus and Prevotella spp. dominant. Further dissection of the microbial composition of the developing lesion will be enhanced by development of methods for enriching the display in population analysis of low abundance species of interest. The sequence information obtained will be used to design specific oligonucleotide probes for the determination of the spatial localization of organisms within the lesion by fluorescence in situ hybridization combined with confocal microscopy. Experimental analysis will extend to provide information on the mechanisms of bacterial invasion of the Dental pulp at high resolution using both extractive and in situ approaches and relate this to the histopathological responses of the pulps. Data obtained from this study will provide an objective basis for an understanding of the extension of the carious process with potential diagnostic value for predicting pulp response including pulpal death and endodontic infection.