The Role of Uncultivated Bacteria in Bacterial Vaginosis

描述（由申请人提供）：人类阴道的微生物群可以深刻影响妇女和新生儿的健康。例如，患有细菌性阴道病（BV）的妇女获得性传播感染（如艾滋病毒）、患盆腔炎和早产的风险增加。与BV相关的医疗保健成本是巨大的，无论是考虑到每年数百万次阴道分泌物的医疗保健访问相关的直接成本，还是间接成本，如早产儿的护理。尽管最近的进展已经使用分子方法（PCR）来更好地描述BV的微生物学，但对于为什么一些妇女会发生这种情况缺乏了解。女性发生BV后阴道微生物群的变化得到了很好的描述，但对BV的微生物，生理和行为前因了解甚少。该提案旨在通过在一项旨在捕获事件和复发病例的纵向研究中收集关于有BV风险的妇女的数据，来克服这一知识差距。将使用广泛的16S rRNA基因PCR结合焦磷酸测序和细菌特异性定量PCR来表征阴道微生物菌群。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，避孕套的使用）有关。研究导致BV的因素对于了解BV的发病机制至关重要。这些知识可用于开发改进的诊断，预防和治疗方法。虽然抗生素如甲硝唑和克林霉素可用于治疗BV，但复发是常见的，3个月时复发率高达30%，1年时复发率>50%。这种高复发率的原因尚不清楚。高复发率使改善BV不良健康后果的努力复杂化。该提案旨在通过研究可能允许阴道播种的细菌的阴道外储库来描述与BV复发相关的微生物因素。将在抗生素治疗期间和治疗后对BV女性（和一部分性伴侣）进行随访，以使用PCR评估阴道和阴道外部位的阴道细菌浓度。预计阴道外储库定植将是BV复发的主要风险因素。生物膜在BV发病机制中是否重要仍有争议，基于一些小型横断面研究的有趣数据支持这一概念。将使用荧光原位杂交（FISH）评估从有和没有BV的女性中获得的阴道样本中细菌生物膜的患病率。本文将研究非活检样本的价值。抗生素治疗后将评估阴道细菌生物膜的变化，以确定BV复发是否与生物膜群落中细菌的持续存在相关。如果生物膜在BV中显示出重要性，这将为基于生物膜形成和细胞信号传导的破坏的治疗和预防开辟许多新途径。

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.