GENETIC PROBE ANALYSIS OF SUBGINGIVAL PLAQUE SPIROCHETES

龈下菌斑螺旋体的基因探针分析

• 批准号: 2131635
• 负责人: Everett P Greenberg
• 金额: $ 14.25万
• 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/2131635
• 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构成了龈下菌斑的很大一部分 与牙周病有关的细菌菌群。 虽然螺旋体 已被认为是牙周病的病因，它们 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涉及困难 培养和计数单个螺旋体物种。 分子 将开发快速定量相关物质的技术 龈下菌斑中单个螺旋体物种的丰度 样品。 这是理解角色的关键一步 牙周病中的螺旋体。 分子方法下 开发涉及耦合 PCR 荧光寡核苷酸探针 分析同时估计几种的相对丰度 个别口腔螺旋体种类、螺旋体总数，以及 龈下菌斑样本中的真细菌总数。 这 整个过程将很快，在不到两天的时间内提供数据 样品采购时间，大量样品可 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可实现灵敏、 高度特异性、同时多物种分析。 这是第一步 旨在设计有意义的临床研究，其中定义明确 牙周病状态或疾病活动度与 特定螺旋体的丰度。 长期目标是利用 PCR-荧光探针程序以开展适当的生态研究 旨在了解口腔螺旋体与牙周的关系 疾病。 此外，预计分析技术 开发可能对其他细菌的研究具有广泛的适用性 天然样品。