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: 6818071
• 负责人: Ann L Griffen
• 金额: $ 44.59万
• 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/6818071
• 关键词: bacteria characteristic; bacterial cytopathogenic effect; bacterial genetics; biofilm; breast feeding; clinical research; dental caries; dental development; dentinogenesis; dentition; health disparity; human subject; infant human (0-1 year); low socioeconomic status; middle childhood (6-11); molecular cloning; nucleic acid quantitation /detection; nucleic acid sequence; oral bacteria; polymerase chain reaction; preschool child (1-5); species difference

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.

龋齿是儿童最常见的慢性疾病，也是美国儿童最大的未满足的医疗保健需求。患病率和治疗率的社会经济差异是一个重大的公共卫生问题。迄今为止，尚未开发出有效的生物干预措施来预防龋齿。牙菌斑包含数百种不同的生物体，其中许多研究很少。许多物种已被证明产生足够的酸驱动pH值低于临界水平，并容忍低pH值的研究主要集中在变形链球菌作为龋齿的病原体，但基于以前的研究人员使用培养和我们的初步研究结果，使用分子方法，S。变形菌并不总是存在于龋齿中，经常以低水平发现，并且额外的和意想不到的细菌种类可能是重要的。此外，相对较少的注意力已经支付给确定健康相关的和潜在的有益的细菌物种，可能驻留在口腔中。在拟议的项目中，将通过克隆和测序从牙菌斑样本分离的DNA扩增的细菌16S核糖体基因来确定儿童龋齿和健康中存在的细菌种类。这种开放式方法将允许检测和鉴定存在的所有细菌物种，包括新的、未培养的或意外的物种。通过这种方法鉴定的与龋齿或健康潜在相关的物种的存在和数量将使用定量实时PCR来确定。这种双技术方法将允许检查比单独克隆和测序可能的大得多的样本量。完成拟议的研究将确定与乳牙和年轻恒牙列严重龋齿的发生和发展相关的病原体，并确定与健康牙列相关的细菌种类。这项研究还将提供一个基于分子技术的儿童龈上植物群的综合目录。拟议的工作的意义在于，识别额外的龋齿病原体将为生物干预提供替代目标，并且识别有益的健康相关物种可以为建立抗龋齿微生物群落的治疗干预提供基础。