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Composition of the Vaginal Microbiota in Women of Reproductive Age - Sensitive and Specific Molecular Diagnosis of Bacterial Vaginosis Is Possible?

机译:育龄妇女阴道菌群的组成-细菌性阴道病的敏感性和特异性分子诊断是否可能?

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摘要

Background and Objective: Bacterial vaginosis (BV) is the most common vaginal disorder, characterized by depletion of the normal lactobacillus-dominant microbiota and overgrowth of commensal anaerobic bacteria. This study aimed to investigate the composition of the vaginal microbiota in women of reproductive age (healthy women and women with BV), with the view of developing molecular criteria for BV diagnosis. Materials and Methods: Vaginal samples from 163 women (79 control, 73 BV and 11 intermediate (Lactobacillary grade II flora) cases) were analyzed using 454 pyrosequencing of the hypervariable regions V3-V4 of the 16S rRNA gene and 16 quantitative bacterial species/genus-specific real-time PCR assays. Sensitivities and specificities of potential BV markers were computed using the Amsel criteria as reference standard for BV. The use of quantitative thresholds for prediction of BV, determined for both relative abundance measured with 454 pyrosequencing and bacterial load measured with qPCR, was evaluated. Results: Relative to the healthy women, the BV patients had in their vaginal microbiota significantly higher prevalence, loads and relative abundances of the majority of BV associated bacteria. However, only Gardnerella vaginalis, Atopobium vaginae, Eggerthella, Prevotella, BVAB2 and Megasphaera type 1 detected at or above optimal thresholds were highly predictable for BV, with the best diagnostic accuracy shown for A. vaginae. The depletion of Lactobacillus species combined with the presence of either G. vaginalis or A. vaginae at diagnostic levels was a highly accurate BV predictor. Conclusions: Quantitative determination of the presence of G. vaginalis, A. vaginae, Eggerthella, Prevotella, BVAB2 and Megasphaera type 1 as well as the depletion of Lactobacillus was highly accurate for BV diagnosis. Measurements of abundance of normal and BV microbiota relative to total bacteria in vaginal fluid may provide more accurate BV diagnosis, and be used for test-of-cure, rather than qualitative detection or absolute counts of BV related microorganisms.
机译:背景与目的:细菌性阴道病(BV)是最常见的阴道疾病,其特征是正常乳杆菌主导的微生物群耗竭,共生厌氧菌过度生长。这项研究旨在调查育龄妇女(健康妇女和BV妇女)阴道微生物群的组成,以期建立BV诊断的分子标准。材料和方法:使用454焦磷酸测序对16S rRNA基因的高变区V3-V4和16种定量细菌种类/属进行分析,从163名妇女(79名对照,73名BV和11名中级(乳杆菌II级菌群)病例)的阴道样本中进行了分析。特定的实时PCR分析。使用Amsel标准作为BV的参考标准,计算潜在BV标记的敏感性和特异性。评估了定量阈值用于BV的预测,该阈值用于确定454焦磷酸测序法测定的相对丰度和qPCR测定的细菌载量。结果:相对于健康女性,BV患者的阴道微生物群中大多数BV相关细菌的患病率,负荷和相对丰度明显更高。但是,只有在最佳阈值或更高的最佳阈值下检测到的阴道加德纳菌,阴道拟南芥,埃奇特氏菌,普雷沃特菌,BVAB2和1型巨噬球菌才可预测BV,而对阴道曲霉的诊断准确性最高。乳酸杆菌物种的耗竭与诊断水平的阴道加德纳氏菌或阴道加曲霉的存在相结合是一种高度准确的BV预测因子。结论:定量测定阴道G.,阴道曲霉,Eggerthella,Prevotella,BVAB2和Megasphaera 1型的存在以及乳酸菌的消耗对于BV诊断是高度准确的。正常和BV菌群相对于阴道液中细菌总数的测量可以提供更准确的BV诊断,并且可以用于治愈测试,而不是BV相关微生物的定性检测或绝对计数。

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