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%的牙周炎可归因于吸烟，吸烟者比不吸烟者患更严重和更广泛疾病的风险更高。虽然细菌菌斑和吸烟都被报道在牙周炎中起重要作用，但个体细菌种类或群体与吸烟之间的关系尚未得到很好的阐明。最近对牙龈下微生物群落的独立培养探索表明，大多数物种是未经培养的，因此在使用传统的基于培养的方法时无法识别。因此，使用分子方法来检查这一高危人群的龈下微生物特征是很重要的，这种方法可以避免培养的需要。我们假设吸烟与牙龈下微生物谱的改变有关，并且这种改变与疾病严重程度无关。我们建议使用指向16S rRNA基因的头状探针，用流式细胞仪同时测量多个物种的水平。具体目标1提出，通过比较牙周健康吸烟者和牙周炎患者之间培养物种和未培养物种的水平，研究吸烟是否与牙龈下生态系统的全球扰动有关。这些物种是根据我们早期使用开放式分子方法进行细菌鉴定的工作选择的。具体目标2将调查吸烟者微生物谱的改变是否可归因于该队列中疾病严重程度的增加。我们将比较牙周炎严重程度相似的当前吸烟者和从不吸烟者之间选定生物体的水平。这些研究首次全面研究了吸烟与龈下微生物群落变化之间的关系，并将为更广泛的研究建议提供重要的初步数据，以评估与高危人群牙周炎发病和进展相关的细菌变化。在本研究过程中，开发出一种灵敏、高通量的定量分析方法，可用于任何微生物生态系统的表征，并将填补急需的方法学空白。在这一高危人群中鉴定假定的病原体和与健康相关的细菌将有助于更好地了解吸烟在牙周病病因学中的作用。这是开发临床有用的疾病标志物和靶向生物干预措施的重要一步。关键词：牙周炎，吸烟，细菌，龈下，16S，分子微生物学，流式细胞术这些研究代表了第一个全面的方法来检查吸烟和牙龈下微生物群落变化之间的关系。在这一高危人群中鉴定假定的病原体和与健康相关的细菌将是研究它们与人类宿主相互作用的重要一步，并可能使人们更好地了解吸烟在牙周病病因学中的作用。这可能会导致临床有用的疾病标志物和预测因素的发展，以及预防疾病的生物干预措施。