Genomic Tools For Studying The Ecology Of The Human Vaginal Microflora

描述(由申请人提供)：健康育龄妇女的正常阴道菌群在防止“不良”微生物成功定居方面起着关键作用，包括细菌性阴道病、酵母菌感染、性传播疾病和尿路感染。我们的长期目标是准确了解正常、健康女性的阴道微生物生态系统的组成和生态，作为了解正常微生物区系如何降低感染这些传染病的风险和确定决定疾病易感性的因素的基本前提。具体的假设是：1.妇女的遗传背景影响阴道微生物群的组成；2.虽然不同妇女的阴道群落组成明显不同，但一个群落的功能潜力是相似的。将开发检验这些假说所需的工具，并通过以下具体目标解决这些工具：1.利用16S rRNA基因测序分析对400名不同种族背景的妇女的阴道微生物进行全面调查。2.开发基于16S rDNA的分子工具(分子倒置探针)，快速、定量地测量阴道微生物群落的微生物种类组成和丰度。3.用群落基因组学方法研究健康女性阴道微生物区系的代谢潜能。我们将对五种最主要的群落类型的“群落基因组”进行排序。序列数据和分析将通过基于网络的阴道微生物组数据库免费获取。4.开发阴道微生物组表达基因芯片(C)阵列，这是一种基于高密度寡核苷酸微阵列的工具，用于阴道微生物群落的功能基因组分析。使用这个工具，我们将在两个月的过程中评估社区基因的表达，同时监测生殖激素水平。我们认为这项工作类似于人类基因组计划，在该计划中，初始序列信息为未来数十年专注于人类健康和疾病的方方面面的工作提供了一个平台。同样，这些工具将对妇女健康的研究和评估至关重要。

DESCRIPTION (provided by applicant): The normal vaginal microflora in healthy women of reproductive age plays a key role in preventing successful colonization by "undesirable" organisms including those responsible for bacterial vaginosis, yeast infections, sexually transmitted diseases and urinary tract infections. Our long-term goal is to develop an accurate understanding of the composition and ecology of the vagina microbial ecosystem in normal, healthy women as an essential prerequisite for comprehending how the normal microflora reduces the risk of acquiring these communicable diseases and for defining the factors determining disease susceptibility. The specific hypotheses are that 1. women's genetic backgrounds influence the composition of the vaginal microflora, 2. while vaginal community compositions markedly vary amongst women, the functional potential of a community is similar. Tools necessary to test these hypotheses will be developed and are addressed by these specific aims: 1. Conduct a comprehensive survey of the vaginal microflora in 400 women of different ethnic background using 16S rRNA gene sequencing analysis. We will correlate the community composition with genetic background, and provide the basis for Aim 2. 2. Develop a 16S rDNA-based molecular tool (molecular inversion probes) to rapidly and quantitatively measure the microbial species composition and abundance of a vaginal community. 3. Characterized the metabolic potential of the vaginal microflora of healthy women by community genomics. We will sequence the "community genome" of the five most predominant community types. Sequence data and analysis will be freely accessible through the web-based Vaginal Microbiome Database. 4. Develop the Vaginal Microbiome Expression GeneChip(c) array, a high density oligonucleotide microarray- based tool for functional genomic analyses of vaginal microbial communities. Using this tool we will assess community gene expression over the course of two months, while monitoring reproductive hormone levels. We view this work as analogous to the human genome project, in which initial sequence information provided a platform for decades of future work focused on all aspect of human health and disease. Similarly, these tools will be critical for the study and evaluation of women's health.