The Role of Uncultivated Bacteria in Bacterial Vaginosis

未培养的细菌在细菌性阴道病中的作用

• 批准号: 7160565
• 负责人: DAVID Neal FREDRICKS
• 金额: $ 55.74万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7160565
• 关键词: Affect; Aftercare; Anaerobic Bacteria; Antibiotic Therapy; Area; Bacteria; Bacterial Vaginosis; Base Sequence; Behavioral; Biological Assay; Cell Count; Clinical; Collaborations; Collection; Communities; Complement; Complex; Condition; Confocal Microscopy; Consensus Sequence; Count; DNA Sequence; Data; Detection; Diagnosis; Diagnostic; Disease; Ecology; Ecosystem; Environment; Epidemiology; Epithelial; Escherichia coli; Female of child bearing age; Fluorescence; Fluorescence Microscopy; Fluorescent in Situ Hybridization; Fred Hutchinson Cancer Research Center; Gardnerella vaginalis; Gastrointestinal tract structure; Genes; Goals; Grant; HIV Infections; Head; Health; Human; In Situ Hybridization; Infection; Knowledge; Laboratories; Lactobacillus; Libraries; Link; Measures; Megasphaera; Methodology; Methods; Microbe; Microbiology; Microscopic; Minority; Mobiluncus curtisii; Molecular; Molecular Diagnostic Techniques; Molecular Epidemiology; Molecular Probes; Mucous Membrane; Natural History; Nucleic acid sequencing; Numbers; Oral cavity; Oral mucous membrane structure; Outcome; Pathogenesis; Patients; Pelvic Inflammatory Disease; Play; Polymerase Chain Reaction; Population; Premature Labor; Prevalence; Prevention; Range; Reagent; Recombinant DNA; Rectum; Relapse; Reproductive Health; Research Infrastructure; Research Personnel; Resistance; Resource Sharing; Ribosomal DNA; Risk Factors; Role; Sampling; Sequence Analysis; Sex Behavior; Sexually Transmitted Diseases; Specimen; Surface; Swab; Tissues; Unsafe Sex; Vagina; Vaginal Discharge; Woman; Women' s Health; bacteria characteristic; cohort; experience; improved; microbial; microbial community; novel; pathogen; prevent; programs; rRNA Genes; rectal; reproductive success; tool; vaginal fluid

DESCRIPTION (provided by applicant): Bacterial vaginosis (BV) is a condition that affects millions of women, produces malodorous vaginal discharge, and is linked to several serious health conditions including preterm labor and HIV infection. The cause of BV is unclear, though changes in vaginal flora are noted and patients may respond to antibiotic treatment. No single cultivated bacterium is specifically associated with BV. Bacteria in complex microbial communities can be identified without cultivation by characterizing ribosomal DNA sequences. This approach offers the advantage of detecting fastidious or cultivation-resistant microbes. The short-term objective of this project is to use cultivation-independent molecular methods to characterize the vaginal flora of women with BV and healthy controls. The long-term objectives are to understand the bacterial ecology of the vaginal niche and determine the role uncultivated bacteria play in BV. Hypothesis: novel communities of bacteria, including many uncultivated species, are found in subjects with BV. To meet these objectives, the following aims will be pursued: Aim 1: Identify the community of bacteria associated with BV. Broad range PCR will be used to amplify bacterial 16S rRNA genes present in vaginal fluid. Sequence analysis of the amplified 16S rDNA will be used to identify the bacterial species found in subjects with BV and healthy controls. Aim 2: Use bacterium specific PCR and fluorescence in situ hybridization (FISH) assays to detect and quantify each vaginal bacterium. Bacterium-specific PCR and FISH assays are more sensitive than broad range PCR for detecting infrequent bacterial species in complex communities. These methods will be applied to vaginal fluid samples from additional subjects with BV and healthy controls. FISH paired with fluorescence microscopy will be used to count bacteria in vaginal fluid smears, thus producing an independent measure of bacterial representation to complement PCR methods. Aim 3: Determine if there is an extra-vaginal niche for BV associated bacteria. The gastrointestinal tract hosts numerous anaerobic bacteria and may be a reservoir for the bacteria linked to BV. Swabs of extra-vaginal mucosal surfaces will be obtained from women with and without BV. Bacteria on these swabs will be detected using bacterium-specific PCRs. We hypothesize that BV-associated bacteria will be common colonizers of the human gastrointestinal tract. Knowledge gained from this project will help identify the bacterial communities found in subjects with BV, will generate opportunities to advance the diagnosis of BV through new PCR and FISH assays, and may help in the prevention of BV by identifying extra-vaginal reservoirs of infection.

描述（由申请人提供）：细菌性阴道病（BV）是一种影响数百万妇女的疾病，产生恶臭的阴道分泌物，并与几种严重的健康状况有关，包括早产和艾滋病毒感染。细菌性阴道炎的病因尚不清楚，但阴道菌群的变化值得注意，患者可能对抗生素治疗有反应。没有一种单独培养的细菌与细菌性阴道炎特异性相关。复杂微生物群落中的细菌可以通过鉴定核糖体DNA序列而无需培养。这种方法提供了检测挑剔或培养耐药微生物的优势。该项目的短期目标是使用不依赖于培养的分子方法来表征患有细菌性阴道炎的妇女和健康对照者的阴道菌群。长期目标是了解阴道生态位的细菌生态，并确定未培养细菌在细菌性阴道炎中的作用。假设：在BV患者身上发现了新的细菌群落，包括许多未培养的物种。为了实现这些目标，将追求以下目标：目标1：确定与细菌性肠胃炎相关的细菌群落。宽范围PCR将用于扩增阴道液中存在的细菌16S rRNA基因。扩增的16S rDNA序列分析将用于鉴定BV患者和健康对照组中发现的细菌种类。目的2：利用细菌特异性PCR和荧光原位杂交（FISH）检测和定量每个阴道细菌。细菌特异性PCR和FISH检测在复杂群落中检测不常见的细菌种类比广谱PCR更敏感。这些方法将应用于其他细菌性阴道炎患者和健康对照者的阴道液样本。FISH与荧光显微镜配对将用于阴道液涂片中的细菌计数，从而产生细菌代表性的独立测量，以补充PCR方法。目的3：确定阴道外是否存在细菌性阴道炎相关细菌的生态位。胃肠道是大量厌氧菌的宿主，可能是与细菌性肠胃炎有关的细菌的储存库。阴道外粘膜表面拭子将从有或没有细菌性阴道炎的妇女身上获得。将使用细菌特异性pcr检测这些拭子上的细菌。我们假设bv相关细菌将是人类胃肠道的常见定植菌。从该项目中获得的知识将有助于确定细菌性阴道炎患者体内发现的细菌群落，将为通过新的PCR和FISH检测来推进细菌性阴道炎的诊断创造机会，并可能通过确定阴道外感染库来帮助预防细菌性阴道炎。