The Role of Uncultivated Bacteria in Bacterial Vaginosis

描述(由申请人提供)：人类阴道的微生物区系可以深刻地影响妇女和新生儿的健康。例如，患有细菌性阴道病(BV)的妇女感染艾滋病毒等性传播疾病、患盆腔炎和早产的风险增加。与BV相关的医疗成本是巨大的，无论是与每年数百万次阴道排液相关的直接成本，还是与早产儿护理等间接成本相关的成本。尽管最近使用分子方法(PCR)来更好地描述BV的微生物学方面取得了进展，但对于一些妇女为什么会患上这种疾病还缺乏了解。女性发生BV后阴道微生物区系的变化已被很好地描述，但对BV的微生物学、生理学和行为学前驱因素了解较少，也缺乏联系。这项建议寻求通过在一项旨在捕捉发病和复发病例的纵向研究中收集BV风险妇女的数据来克服这一知识差距。16S rRNA基因扩增结合焦磷酸测序和细菌特异性定量聚合酶链式反应将用于阴道微生物的鉴定。阴道细菌群落的变化将与生理因素(月经)、药物使用(激素避孕药、抗生素)和行为因素(性行为、避孕套使用)有关。研究导致BV的因素对于了解BV的发病机制至关重要。这些知识可以用来开发改进的诊断、预防和治疗方法。虽然甲硝唑和克林霉素等抗生素可以用来治疗BV，但复发很常见，3个月复发率高达30%，一年复发率高达50%。复发率如此之高的原因尚不清楚。高复发率使减轻BV不良健康后果的努力复杂化。这项建议试图通过研究阴道外的细菌储存库来描述与BV复发相关的微生物因素，这些细菌可能允许阴道播种。在抗生素治疗期间和之后，将对患有BV的妇女(以及部分性伴)进行跟踪，以使用聚合酶链式反应评估阴道和阴道外部位的阴道细菌浓度。预计阴道外蓄水池的定植将是BV复发的主要危险因素。生物膜在BV的发病机制中是否重要仍在争论中，一些基于一些小型横断面研究的有趣数据支持了这一概念。荧光原位杂交(FISH)将被用来评估从患有和不患有BV的妇女身上获得的阴道样本中细菌生物膜的流行率。这里将研究非活组织检查样本的价值。在抗生素治疗后，将评估阴道细菌生物膜的变化，以确定BV的复发是否与细菌在生物膜群落中的持久性有关。如果生物膜在BV中被证明是重要的，这将为基于破坏生物膜形成和细胞信号的治疗和预防开辟许多新的途径。

DESCRIPTION (provided by applicant): The microbiota of the human vagina can profoundly affect the health of women and neonates. For instance, women with the condition bacterial vaginosis (BV) have increased risks of acquiring sexually transmitted infections such as HIV, developing pelvic inflammatory disease, and experiencing preterm birth. Healthcare costs associated with BV are enormous, whether considering the direct costs related to the millions of healthcare visits annually for vaginal discharge, or the indirect costs such as care of pre-term infants. Despite recent advances that have used molecular methods (PCR) to better describe the microbiology of BV, there is a lack of understanding regarding why some women develop this condition. Changes in the vaginal microbiota after women develop BV are well described, but the microbiological, physiological, and behavioral antecedents to BV are poorly understood and connected. This proposal seeks to overcome this knowledge gap by collecting data on women at risk for BV in a longitudinal study designed to capture incident and relapsing cases. Broad range 16S rRNA gene PCR with pyrosequencing and bacterium-specific quantitative PCR will be used to characterize vaginal microbiotas. Changes in vaginal bacterial communities will be linked to physiological factors (menses), medication use (hormonal contraceptives, antibiotics), and behavioral factors (sexual practices, condom use). Studying the factors that lead to BV is critical for understanding pathogenesis. This knowledge can be used to develop improved methods for diagnosis, prevention, and treatment. Although antibiotics such as metronidazole and clindamycin can be used to treat BV, recurrence is common, with recurrence rates of up to 30% at 3 months and >50% at one year. The reasons for this high rate of recurrence are not known. High relapse rates complicate efforts to ameliorate the adverse health consequences of BV. This proposal seeks to describe the microbiological factors associated with recurrence of BV by studying extra-vaginal reservoirs of bacteria that may allow seeding of the vagina. Women with BV (and a subset of sex partners) will be followed during and after antibiotic treatment to assess concentrations of vaginal bacteria in vaginal and extra-vaginal sites using PCR. It is anticipated that colonization of extra-vaginal reservoirs will be a major risk factor for recurrence of BV. There is ongoing debate whether biofilms are important in the pathogenesis of BV, with intriguing data supporting this concept based on some small cross-sectional studies. Fluorescence in situ hybridization (FISH) will be used to assess the prevalence of bacterial biofilms in vaginal samples obtained from women with and without BV. The value of non-biopsy samples will be studied here. Changes in vaginal bacterial biofilms will be assessed after antibiotic treatment to determine if recurrence of BV is associated with persistence of bacteria in biofilm communities. If biofilms are shown to be important in BV, this would open many new avenues for treatment and prevention based on disruption of biofilm formation and cell signaling.