GENETIC PROBE ANALYSIS OF SUBGINGIVAL PLAQUE SPIROCHETES

螺旋体作为一个类群，构成了龈下菌斑的一大部分。 与牙周病有关的细菌群。虽然螺旋体 都被认为是牙周疾病的病原体，他们的 在这些疾病中的作用尚不清楚。知识的匮乏 关于螺旋体在口腔生态学中的作用涉及到困难 在培养和计数单个螺旋体物种方面。分子 将开发技术来快速量化相对 龈下菌斑中单个螺旋体物种的丰度 样本。这是朝着理解 牙周病中的螺旋体。从分子水平上看 研究涉及一种耦合的聚合酶链式反应-荧光寡核苷酸探针 多组分相对丰度的同时估计分析 单个口腔螺旋体物种，总螺旋体种群，以及 龈下菌斑样本中的总细菌数量。这个 整个过程将是快速的，在不到两天的时间内从 样品采购的时间，以及大量的样品可以 同时处理。聚合酶链式反应技术与自动化的结合 荧光寡核苷酸分析技术允许一种灵敏、 高度特异、同时进行的多物种分析。这是第一步 朝着设计有意义的临床研究迈进，在这些研究中 牙周疾病状态或疾病活动性与 大量的特定螺旋体。长期目标是使用 聚合酶链式反应-荧光探针法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，预计分析技术将在 这一发展可能对其他细菌的研究具有广泛的适用性 天然样品。

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.