GENETIC PROBE ANALYSIS OF SUBGINGIVAL PLAQUE SPIROCHETES

龈下菌斑螺旋体的基因探针分析

• 批准号: 2331325
• 负责人: Everett P Greenberg
• 金额: $ 15.28万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-02-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2331325
• 关键词: Bacteroides gingivalis; Treponema; bacterial DNA; dental plaque; fluorescence microscopy; fluorescent dye /probe; gingivitis; method development; microorganism classification; nucleic acid probes; nucleic acid sequence; oligonucleotides; oral bacteria; periodontitis; polymerase chain reaction; ribosomal DNA; ribosomal RNA; species difference

Spirochetes as a group comprise a large portion of the subgingival plaque bacterial flora involved in periodontal diseases. Although spirochetes have been implicated as etiologic agents in periodontal diseases, their role in these diseases is not understood. The lack of knowledge concerning the role of spirochetes in oral ecology relates to difficulty in culturing and enumerating individual spirochete species. Molecular techniques will be developed for rapid quantitation of the relative abundances of individual spirochete species from subgingival plaque samples. This is a critical step towards understanding the roles of spirochetes in periodontal diseases. The molecular approach under development involves a coupled PCR-fluorescent oligonucleotide probe analysis to simultaneously estimate the relative abundances of several individual oral spirochete species, the total spirochete population, and the total eubacterial population in a subgingival plaque sample. The overall procedure will be rapid, providing data in less than two days from the time of sample procurement, and large numbers of samples can be processed simultaneously. The coupling of PCR technology and automated fluorescent oligonucleotide analytical technology allows for a sensitive, highly specific, simultaneous-multispecies analysis. This is a first step towards designing meaningful clinical studies where well-defined periodontal disease states or disease activity are correlated with abundances of specific spirochetes. The long-term objective is to use the PCR-fluorescent probe procedure to develop appropriate ecological studies aimed at understanding oral spirochetes as they relate to periodontal diseases. Also, it is anticipated that the analytical technique under development may have wide applicability to studies of other bacteria in natural samples.

螺旋体作为一个群体包括龈下菌斑的大部分 牙周病涉及的细菌植物群。 虽然螺旋体 作为牙周病的病原体， 在这些疾病中的作用尚不清楚。 缺乏知识 关于螺旋体在口腔生态学中的作用， 培养和计数单个螺旋体物种。 分子 将开发快速定量相对 龈下菌斑中单个螺旋体的丰度 样品 这是理解以下角色的关键一步： 牙周病的症状有哪些 分子方法 开发涉及偶联PCR-荧光寡核苷酸探针 分析，同时估计几个相对丰度 个体口腔螺旋体种类、总螺旋体种群，以及 龈下菌斑样本中的总真细菌群。 的 整个程序将是迅速的，在不到两天的时间内提供数据， 样品采购的时间，以及大量的样品可以 同时处理。 PCR技术与自动化技术的结合 荧光寡核苷酸分析技术允许灵敏， 高度特异性的多物种分析 这是第一步 设计有意义的临床研究， 牙周疾病状态或疾病活动与 丰富的特定螺旋体。 长期目标是利用 PCR-荧光探针程序，以开展适当的生态学研究 旨在了解口腔螺旋体，因为它们与牙周炎有关， 疾病 此外，预计分析技术将在 开发可能具有广泛的适用性，以研究其他细菌， 天然样品