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）：该提案的目的是开发一种新颖的组合标记和检测技术，以成像和区分大多数人类口腔微生物界的成员。 16S rRNA序列的荧光原位杂交（FISH）将在视觉上将微生物群落的个别成员视为收集或原位。通过采用荧光报告基团的组合来创建“光谱特征”，可以扩展鱼类的能力，从而可以同时编码和成像数十个至数百种不同的微生物系统型。增强的，最先进的光谱成像技术将记录光谱特征。新型算法将开发以解构标记的微生物的光谱特征。这种新的成像能力将允许人类口腔微生物群落中的系统型全面普查，并确定该社区的空间组织。原则上，可以通过与16S rRNA的探针同时标记微生物进行系统发育鉴定和针对mRNA或蛋白质的探针来扩展组合标记策略，以进行代谢或结构信息。评估特定微生物的相对丰度，组织和功能的能力将为对口腔微生物群落的生态学进行全面和详细研究开放新的途径。口腔微生物与龋齿，牙龈炎和牙周炎等疾病有关，这些疾病是人类中最常见的细菌感染之一，以及包括心血管疾病，肺部感染以及低出生症和预期妊娠结局在内的全身性疾病。没有任何单一的微生物物种被认为是这些疾病的病因学药，而是假设社区中多种微生物系统型的复杂关联负责维持健康和引起疾病。这里提出的技术的开发将使人们对正常的人类口腔菌群有全面的了解，并促进研究，监测和控制感染的新有效策略。