Molecular Analysis of the Biofilm in Caries and Health

龋齿与健康中生物膜的分子分析

• 批准号: 7232407
• 负责人: Ann L Griffen
• 金额: $ 42.66万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7232407
• 关键词: Acids; Americas; Attention; Biological; Biological Assay; Cataloging; Catalogs; Characteristics; Child; Childhood; Chronic Disease; Cloning; DNA; Data; Dental Plaque; Dental caries; Dentition; Depth; Detection; Development; Disease; Experimental Designs; Genes; Goals; Health; Healthcare; Intervention; Investigation; Lesion; Methods; Microbial Biofilms; Molecular; Molecular Analysis; Numbers; Oral cavity; Organism; Pathogenesis; Patients; Polymerase Chain Reaction; Population; Prevention; Primary Dentition; Public Health; Rate; Research; Research Personnel; Resistance; Risk; Sample Size; Sampling; Staging; Streptococcus mutans; Techniques; Technology; Therapeutic Intervention; Time; Tooth structure; Work; base; microbial; microbial community; microorganism; novel; pathogen; permanent tooth; prevent; socioeconomics

DESCRIPTION: Dental caries is the most common chronic disease of childhood, and is the biggest unmet health care need among America's children. Socioeconomic disparities in both rates of disease and treatment are a major public health issue. To date, effective biological interventions to prevent caries have not been developed. Dental plaque contains several hundred different organisms, many of which are poorly studied. A number of species have been shown to produce sufficient acid to drive pH below critical levels and to tolerate low pH. Research has primarily focused on Streptococcus mutans as the etiologic agent in caries, but based the work of previous investigators using cultivation and on our preliminary findings using molecular methods, S. mutans is not always present in caries, is often found at low levels, and additional and unexpected bacterial species may be important. In addition, comparatively little attention has been paid to identifying health-associated and potentially beneficial bacterial species that may reside in the oral cavity. For the proposed project, bacterial species present in childhood caries and health will be identified by cloning and sequencing bacterial 16S ribosomal genes amplified from DNA isolated from plaque samples. This open ended approach will allow the detection and identification of all bacterial species present, including novel, uncultivated or unexpected species. The presence and quantities of the species identified by this approach as potentially associated with caries or health will then be determined using quantitative, real-time PCR. This dual-technique approach will allow the examination of a much larger sample size than is possible by cloning and sequencing alone. Accomplishment of the proposed studies will identify the pathogens associated with the onset and progression of severe caries of the primary and young permanent dentition, and identify those bacterial species associated with a healthy dentition. This study will also provide a comprehensive catalog of the supragingival flora in children based on molecular technology. The significance of the proposed work is that identification of additional caries pathogens would provide alternative targets for biological interventions, and identification of beneficial health-associated species could provide the basis for therapeutic interventions to establish caries-resistant microbial communities.

描述：龋齿是儿童最常见的慢性疾病，也是美国儿童中最大的未得到满足的医疗保健需求。发病率和治疗率方面的社会经济差距是一个重大的公共卫生问题。到目前为止，还没有开发出有效的预防龋病的生物干预措施。牙菌斑包含数百种不同的微生物，其中许多生物的研究很少。许多物种已被证明能产生足够的酸，使pH低于临界水平，并能耐受低pH。研究主要集中在变形链球菌作为龋病病原体，但基于以前研究人员使用培养的工作和我们使用分子方法的初步发现，变形链球菌并不总是存在于龋齿中，经常在低水平发现，额外的和意想不到的细菌物种可能是重要的。此外，对识别可能存在于口腔中的与健康相关的和潜在有益的细菌种类的关注相对较少。在拟议的项目中，通过克隆和测序从菌斑样本中提取的DNA扩增的细菌16S核糖体基因，将鉴定儿童龋齿和健康中存在的细菌物种。这种开放式的方法将允许检测和鉴定所有存在的细菌物种，包括新的、未培养的或意想不到的物种。通过这种方法确定的可能与龋齿或健康相关的物种的存在和数量将使用定量、实时聚合酶链式反应来确定。这种双重技术方法将允许检验比单独克隆和测序可能的样本量大得多的样本量。完成拟议的研究将确定与乳牙和年轻恒牙严重龋齿的发生和发展有关的病原体，并确定与健康牙列有关的细菌种类。这项研究还将提供基于分子技术的儿童牙龈上菌群的全面目录。拟议工作的意义在于，识别更多的龋病病原体将为生物干预提供替代目标，而与健康相关的有益物种的识别可以为建立抗龋性微生物群落的治疗干预提供基础。