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: 6917254
• 负责人: Ann L Griffen
• 金额: $ 42.5万
• 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/6917254
• 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.

描述：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。发病率和治疗方面的社会经济差异是一个主要的公共卫生问题。迄今为止，预防龋齿的有效生物干预措施尚未开发出来。牙菌斑含有几百种不同的微生物，其中许多还没有得到充分的研究。许多物种已被证明能够产生足够的酸，使pH值低于临界水平，并耐受低pH值。研究主要集中在变形链球菌作为龋齿的病原，但基于先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，通常在低水平上被发现，额外的和意想不到的细菌种类可能很重要。此外，相对而言，人们很少注意识别可能存在于口腔中的与健康相关和潜在有益的细菌种类。对于拟议的项目，将通过克隆和测序从菌斑样本中分离的DNA扩增的细菌16S核糖体基因来鉴定儿童龋齿和健康中存在的细菌种类。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，然后将使用定量实时PCR确定。这种双技术方法将允许检查比单独克隆和测序更大的样本量。完成拟议的研究将确定与初级和年轻恒牙严重龋病的发病和进展相关的病原体，并确定与健康牙齿相关的细菌种类。本研究还将提供基于分子技术的儿童龈上菌群的全面目录。这项工作的意义在于，鉴定额外的龋齿病原体将为生物干预提供替代靶点，鉴定有益的健康相关物种可以为治疗性干预提供基础，以建立耐龋齿微生物群落。