Quantitative analysis of subgingival bacteria in smokers

吸烟者龈下细菌的定量分析

• 批准号: 7531352
• 负责人: Purnima Kumar
• 金额: $ 11.25万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7531352
• 关键词: Bacteria; Biological; Biological Assay; Data; Depth; Development; Disease; Disease Marker; Ecosystem; Epidemiologic Studies; Etiology; Flow Cytometry; Health; Health Status; Human; Individual; Infection; Intervention; Lead; Measures; Microbiology; Molecular; Numbers; Organism; Periodontal Diseases; Periodontitis; Personal Satisfaction; Play; Population; Prevention intervention; Reporting; Research Design; Research Proposals; Risk; Role; Sampling; Secondary to; Severities; Severity of illness; Smoke; Smoker; Smoking; Tooth Loss; Tooth structure; Widespread Disease; Work; base; cohort; disorder prevention; microbial; microbial community; non-smoker; pathogen; rRNA Genes; tool

DESCRIPTION (provided by applicant): 41.9% of periodontitis in the USA is attributable to smoking, and smokers are at a higher risk for more severe and extensive disease than non-smokers. Although both bacterial plaque and smoking are reported to play important roles in periodontitis, associations between individual bacterial species or consortia and smoking have not been well elucidated. Recent cultivation-independent explorations of the subgingival microbial community have revealed that the majority of species are uncultivated and hence, unrecognized when using traditional, cultivation-based approaches. Therefore, it is important to examine the subgingival microbial profile of this high-risk population using molecular approaches that circumvent the need for cultivation. We hypothesize that smoking is associated with an alteration in the subgingival microbial profile and that this alteration is independent of disease severity. We propose to use bead-probes directed to the 16S rRNA genes to simultaneously measure the levels of multiple species with a flow cytometer. Specific aim 1 proposes to investigate if smoking is associated with a global perturbation of the subgingival ecosystem by comparing levels of cultivated species and as-yet-uncultivated phylotypes between periodontally healthy smokers and those with periodontitis. These species have been selected based on our earlier work using an open-ended molecular approach for bacterial identification. Specific aim 2 will investigate if altered microbial profiles in smokers could be attributed to increased severity of disease in this cohort. We will compare the levels of selected organisms between current and never smokers with similar severity of periodontitis. These studies represent the first comprehensive approach to examine the association between smoking and changes in the subgingival microbial community and will provide important preliminary data for much broader research proposals to evaluate the bacterial changes associated with the onset and progression of periodontitis in this high-risk population. The development of a sensitive, high-throughput quantitative assay during the course of this study can be used in the characterization of any microbial ecosystem and will fill a much-needed methodological niche. Identification of putative pathogens and health-associated bacteria in this high-risk population will lead to a better understanding of the role of smoking in the etiology of periodontal diseases. This is an important step in developing clinically useful disease markers and targeted biological interventions. Keywords: periodontitis, smoking, bacteria, subgingival, 16S, molecular microbiology, flow cytometry. These studies represent the first comprehensive approach to examine the association between smoking and changes in the subgingival microbial community. Identification of putative pathogens and health-associated bacteria in this high-risk population will be an important step in studying their interactions with the human host and could lead to a better understanding of the role of smoking in the etiology of periodontal diseases. This could lead to the development of clinically useful disease markers and predictors as well as biological interventions for prevention of disease.

描述（由申请人提供）：美国41.9％的牙周炎归因于吸烟，与非吸烟者相比，吸烟者患更严重和更广泛的疾病的风险更高。尽管据报道细菌斑块和吸烟在牙周炎中起重要作用，但单个细菌或财团与吸烟之间的关联尚未得到很好的阐明。近似微生物群落的最新培养探索表明，大多数物种是未经文化的，因此在使用基于传统的，基于种植的方法时未被认可。因此，重要的是，使用分子方法来检查这种高风险种群的尺寸微生物谱，以规避培养的需求。我们假设吸烟与尺寸微生物特征的改变有关，并且这种改变与疾病的严重程度无关。我们建议使用针对16S rRNA基因的珠探针同时测量具有流式细胞仪的多种物种的水平。具体目标1提议通过比较牙周健康吸烟者与患有牙周炎的人之间的培养物种水平和尚未分类的系统型，研究吸烟是否与亚生生态系统的全球扰动有关。这些物种是基于我们的早期工作，使用开放式分子方法进行细菌鉴定选择。具体目标2将调查吸烟者中的微生物特征是否会归因于该队列中疾病严重程度的增加。我们将比较当前和从未吸烟的牙周炎严重程度的吸烟者之间的选择水平。这些研究代表了检验吸烟与尺寸微生物群落变化之间关联的第一种综合方法，并将为更广泛的研究建议提供重要的初步数据，以评估这个高风险人群中与牙周炎的发作和进展相关的细菌变化。在这项研究过程中，可以使用敏感，高通量定量测定法的发展，可用于表征任何微生物生态系统，并填充急需的方法论利基市场。在这种高风险人群中鉴定假定的病原体和与健康相关的细菌将使人们更好地了解吸烟在牙周疾病病因中的作用。这是开发临床上有用的疾病标记和靶向生物学干预措施的重要步骤。关键词：牙周炎，吸烟，细菌，尺寸，16S，分子微生物学，流式细胞术。这些研究代表了检查吸烟与次生微生物群落变化之间关联的第一种全面方法。在这种高危人群中鉴定推定的病原体和与健康相关的细菌将是研究其与人类宿主相互作用的重要一步，并可能更好地了解吸烟在牙周疾病病因中的作用。这可能导致临床上有用的疾病标志物和预测因子以及预防疾病的生物学干预措施的发展。