Molecular Analysis of Infection in Advanced Caries

产品说明：龋损延伸到牙本质基质中破坏牙齿的物理和功能完整性，并对下面的牙髓组织提供越来越有害的损害。与这一过程相关的细菌还不太清楚。目前，没有可靠的手段来识别龋损下的牙髓组织的状态，也没有增强的治疗方式可用。这严重损害了目前对这种临床上重要的疾病的诊断和治疗制度的效力。 初步的研究已经使用了一个通用的细菌扩增子的基础上16S rDNA，以确定主要的细菌种群的龋牙本质。这项工作的补充是实时聚合酶链反应定量的细菌DNA，以确定水平的细菌物种的系统发育分析。一个复杂的阵列，主要是厌氧细菌被证明与特征乳杆菌和普雷沃氏菌属。占主导地位。进一步解剖的微生物组成的发展中的病变，将加强开发的方法，丰富显示在人口分析的低丰度物种的利益。所获得的序列信息将用于设计特异性寡核苷酸探针，以通过荧光原位杂交结合共聚焦显微镜来确定病变内生物体的空间定位。 实验分析将扩展到提供信息的细菌入侵的牙髓在高分辨率使用提取和原位方法的机制，并将其与牙髓的组织病理学反应。从这项研究中获得的数据将提供一个客观的基础，了解龋过程的扩展与潜在的诊断价值，预测牙髓反应，包括牙髓死亡和牙髓感染。

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.