New Technologies to Identify Novel Periodontal Pathogens

识别新型牙周病原体的新技术

• 批准号: 8094091
• 负责人: Flavia Rocha Teles
• 金额: $ 14.67万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8094091
• 关键词: Adult; Affect; American; Bacteria; Clinical; Communicable Diseases; Complex; Development; Diagnosis; Disease; Environment; Etiology; Food; Fostering; Future; Goals; Growth; Growth Factor; Health; In Situ; In Vitro; Indigenous; Individual; Infection; Investigation; Knowledge; Laboratories; Lead; Link; Liquid substance; Microbial Biofilms; Minority; Mission; Molecular; Names; Oligonucleotide Probes; Oligonucleotides; Oral; Oral health; Outcome; Pathogenesis; Periodontal Diseases; Periodontitis; Play; Population; Positioning Attribute; Prediabetes syndrome; Prevalence; Prevention; Public Health; RNA; Reporting; Research; Research Personnel; Resources; Ribosomal RNA; Role; Sampling; Screening procedure; Severities; Signaling Molecule; Taxon; Techniques; Technology; Testing; Time; Tooth Loss; Treatment Cost; Virulence; Work; base; bone loss; disorder prevention; distraction; follow-up; functional genomics; improved; innovation; innovative technologies; microbial; new technology; novel; oral bacteria; oral infection; pathogen; subgingival biofilm; tool

DESCRIPTION (provided by applicant): Periodontal diseases affect 35% of US adults and lead to bone and tooth loss. Treatment costs reach U$ 4.4 billion per year to American taxpayers. Studies have linked periodontal diseases and systemic conditions. Over half of the oral taxa have not been cultivated in vitro. There is a gap in knowledge regarding which of those taxa are common and numerous-and therefore relevant - in periodontal health and disease. Our long term goal is to unveil novel periodontal pathogens and determine the mechanisms they use for periodontal breakdown. The overall objective of this proposal is to identify prominent subgingival taxa associated with periodontal health or disease that are currently uncultivated or cultivable but currently unrecognized. Reports of uncultivated/ unrecognized taxa present in periodontitis have recently emerged. Our central hypothesis is that a subset of the uncultivated/unrecognized taxa in subgingival biofilms is pathogenic. Many oral uncultivated/unrecognized taxa are transients and brought in by food or liquids and have little, if any, relevance to periodontal health or disease. Thus, we do not aim at recovering just any "domesticateable" uncultivated subgingival isolate; neither at growing uncultivated taxa in multispecies complex. We aim at identifying numerically prominent uncultivated/unrecognized taxa in periodontal health or disease and recover them in pure culture as a first step in determining their role in periodontal health or disease. The rationale of this proposal is to eliminate the "distraction" represented by transient taxa and focus the search for novel prominent subgingival bacteria. We will use innovative technologies to quantify a wide range of uncultivable/unrecognized taxa in large numbers of biofilm samples. We will test the central hypothesis and accomplish the objective of this application by pursuing the following Specific Aims. 1) We will identify prominent uncultivated/unrecognized bacteria in periodontal health and disease. This will be achieved by screening 320 individual subgingival biofilm samples from 16 periodontally healthy and 16 periodontitis subjects. We will employ an innovative technique developed by us, the RNA-Oligonucleotide Quantification Technique (ROQT). It is based on 16s rRNA targeted oligonucleotide probes and it will determine which of 140 uncultivated/unrecognized species/taxa/ clones are common and numerous in subgingival biofilms. Once prominent taxa are defined, 2) we will use the appropriate probes to isolate in pure culture, cultivable but as yet unrecognized prominent taxa. We will use 15 conditions for growth of those taxa. This aim will distinguish taxa that can be cultivated under normal laboratory conditions. This research is significant because biologically meaningful taxa currently considered to be uncultivated/unrecognized will be isolated, characterized and properly classified. This is crucial for virulence, antigenicity and functional genomics studies and investigation of growth factors and signaling molecules. Results from this study will support NIDCR's mission by leading to the identification of pathogenic mechanisms and novel targets for disease prevention, diagnosis and therapy. PUBLIC HEALTH RELEVANCE: The proposed research is relevant of public health because, despite increasing knowledge of the development of periodontal diseases, these infections affect more than 30% of US adults and are more prevalent among minorities. These diseases have been linked to systemic conditions and their treatment costs billions annually. This project is relevant to NIDCR's mission because understanding of the role of uncultivated/unrecognized subgingival bacteria can provide new targets for prevention, diagnosis and treatment of oral infections.

描述（由申请人提供）：牙周病影响35%的美国成年人，并导致骨骼和牙齿脱落。美国纳税人每年的治疗费用达到44亿美元。研究表明牙周病和全身状况有关。超过一半的口腔分类群没有在体外培养。关于哪些分类群在牙周健康和疾病中是常见的和大量的，因此是相关的，在知识上存在差距。我们的长期目标是揭示新的牙周病原体，并确定它们用于牙周破坏的机制。这项建议的总体目标是确定突出的龈下类群与牙周健康或疾病，目前未培养或可培养，但目前尚未认识到。最近出现了未培养/未识别的分类群存在于牙周炎的报告。我们的中心假设是，龈下生物膜中未培养/未识别的类群的一个子集是致病的。许多口腔未培养/未识别的分类群是短暂的，由食物或液体带入，与牙周健康或疾病几乎没有关系。因此，我们的目标不是恢复任何“驯化”的未栽培的龈下分离，也不是在多物种复杂的生长未栽培的类群。我们的目标是确定在牙周健康或疾病的数量突出的未培养/未识别的类群，并恢复他们在纯培养的第一步，在确定其在牙周健康或疾病的作用。该建议的基本原理是消除以瞬时类群为代表的“分心”，并集中寻找新的突出龈下细菌。我们将使用创新技术来量化大量生物膜样品中的各种不可培养/未识别的类群。我们将通过追求以下具体目标来检验中心假设并实现本申请的目标。1)我们将确定牙周健康和疾病中突出的未培养/未识别的细菌。这将通过筛选来自16名牙周健康和16名牙周炎受试者的320个个体龈下生物膜样本来实现。我们将采用我们开发的创新技术，RNA寡核苷酸定量技术（ROQT）。它是基于16 s rRNA靶向寡核苷酸探针，它将确定哪些140个未培养/未识别的物种/分类群/克隆是常见的和众多的龈下生物膜。一旦确定了突出的分类群，2）我们将使用适当的探针在纯培养物中分离可培养但尚未识别的突出分类群。我们将使用15种条件来生长这些分类群。这一目标将区分分类群，可以在正常的实验室条件下培养。这项研究是重要的，因为生物学上有意义的类群目前被认为是未培养/未识别的将被隔离，特点和适当的分类。这对于毒力、抗原性和功能基因组学研究以及生长因子和信号分子的研究至关重要。这项研究的结果将通过识别致病机制和疾病预防、诊断和治疗的新靶点来支持NIDCR的使命。 公共卫生关系：拟议的研究与公共卫生有关，因为尽管对牙周病发展的了解越来越多，但这些感染影响了超过30%的美国成年人，并且在少数民族中更为普遍。这些疾病与全身性疾病有关，每年的治疗费用高达数十亿美元。该项目与NIDCR的使命相关，因为了解未培养/未识别的龈下细菌的作用可以为预防、诊断和治疗口腔感染提供新的目标。