A Novel Technology to Image Human Oral Microbial Communities

人类口腔微生物群落成像新技术

• 批准号: 7612321
• 负责人: Alex M Valm
• 金额: $ 2.76万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-29 至 2011-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7612321
• 关键词: Algorithms; Arts; Bacteria; Bacterial Infections; Biological Assay; Cardiovascular Diseases; Censuses; Communities; Complex; Data; Data Analyses; Dental caries; Detection; Development; Disease; Ecology; Fluorescence; Fluorescence Microscopy; Fluorescent in Situ Hybridization; Fluorochrome; Fostering; Gingivitis; Goals; Health; Human; Image; Imagery; Imaging Techniques; Imaging technology; In Situ; In Situ Hybridization; Individual; Infection; Infection Control; Label; Low Birth Weight Infant; Lung; Messenger RNA; Metabolic; Methods; Microbe; Molecular; Monitor; Numbers; Oligonucleotide Probes; Oral; Oral cavity; Periodontitis; Phylogenetic Analysis; Pregnancy Outcome; Proteins; Protocols documentation; Relative (related person); Reporter; Research; Ribosomal RNA; Systemic disease; Taxon; Techniques; assay development; combinatorial; community organizations; design; image processing; member; microbial; microbial community; new technology; novel; oral bacteria; oral biofilm; research study; technology development; tool

DESCRIPTION (provided by applicant): The objective of this proposal is to develop a novel combinatorial labeling and detection technology for imaging and distinguishing between most if not all of the members of the human oral microbiological community. Fluorescence In Situ Hybridization (FISH) for 16S rRNA sequences will visually identify individual members of the microbial community either as collected or in situ. The capabilities of FISH will be extended by employing a combination of fluorescent reporter groups to create "spectral signatures" that allow simultaneous encoding and imaging of tens to hundreds of different microbial phylotypes. Enhanced, state-of-the-art spectral imaging techniques will record the spectral signatures. Novel algorithms will be developed to deconstruct spectral signatures of labeled microbes. This new imaging capability will allow a comprehensive census of phylotypes in the human oral microbial community as well as determination of the spatial organization of this community. In principle, the combinatorial labeling strategy could be extended by simultaneously labeling microbes with probes against 16S rRNA for phylogenetic identification and probes against mRNA or proteins for metabolic or structural information. The capability to assess the relative abundance, organization and function of particular microbes will open new avenues for comprehensive and detailed study of the ecology of the oral microbial community. Oral microbes are implicated in diseases such as caries, gingivitis, and periodontitis, which are among the most common bacterial infections in humans, as well as systemic diseases including cardiovascular disease, pulmonary infections, and low birth-weight and pre-term pregnancy outcome. No single microbial species is implicated as the etiological agent of these diseases, rather, it is hypothesized that the complex association of multiple microbial phylotypes in communities are responsible for maintaining health and causing disease. Development of the technology proposed here will allow a comprehensive understanding of the normal human oral flora and foster the development of new and effective strategies to study, monitor, and control infection.

描述(由申请人提供)：本提案的目标是开发一种新的组合标记和检测技术，用于成像和区分大多数(如果不是全部)人类口腔微生物群落的成员。16S rRNA序列的荧光原位杂交(FISH)将直观地识别微生物群落的个别成员，无论是收集的还是原位的。FISH的能力将通过使用荧光报告组的组合来创建“光谱特征”来扩展，该“光谱特征”允许同时编码和成像数十到数百种不同的微生物系统类型。增强的、最先进的光谱成像技术将记录光谱特征。将开发新的算法来解构标记微生物的光谱特征。这一新的成像能力将允许对人类口腔微生物群落的系统类型进行全面普查，并确定该群落的空间组织。原则上，组合标记策略可以扩展为同时用针对16S rRNA的探针标记微生物以鉴定系统发育，并用针对mRNA或蛋白质的探针标记代谢或结构信息。评估特定微生物的相对丰度、组织和功能的能力将为全面和详细研究口腔微生物群落的生态学开辟新的途径。口腔微生物与龋病、牙周炎和牙周炎等疾病有关，这些疾病是人类最常见的细菌感染之一，也与心血管疾病、肺部感染、低出生体重和早孕结局等全身疾病有关。没有单一的微生物物种被认为是这些疾病的病原体，相反，它被假设为群落中多种微生物系统类型的复杂关联负责维持健康和导致疾病。这里提出的技术的发展将使人们能够全面了解正常的人类口腔菌群，并促进新的有效战略的发展，以研究、监测和控制感染。