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Vaginal Microbiota Evaluation and Lactobacilli Quantification by qPCR in Pregnant and Non-pregnant Women: A Pilot Study.
Frontiers in Cellular and Infection Microbiology ( IF 4.6 ) Pub Date : 2020-05-20 , DOI: 10.3389/fcimb.2020.00303
David Pacha-Herrera 1 , Gabriela Vasco 1, 2 , Cecilia Cruz-Betancourt 3 , Juan Miguel Galarza 3 , Verónica Barragán 1 , António Machado 1
Affiliation  

Pregnancy outcomes and women's health are directly affected by vaginal microbiota. This microbiota consists of a dynamic ecosystem of various microbes in different ratios, which in healthy conditions protect the vaginal epithelium from infections. However, cases of vaginal infection are regularly diagnosed in women of reproductive age, contributing to more severe outcomes. Therefore, our main goal was to determine the prevalence of bacterial vaginosis (BV), aerobic vaginitis (AV), and vulvovaginal candidiasis (VVC) among Ecuadorian pregnant and non-pregnant women. A cross-sectional study was conducted among 217 women between 13 and 40 years old seeking primary healthcare in Carlos Andrade Marin Hospital (HCAM), Gynecological-Obstetric Hospital Isidro Ayora (HGOIA) and Center for Teaching Health Cipriana Dueñas during October 2018 to February 2019. The classical characterization of the vaginal microbiota was performed through microscopy by the Nugent criteria to evaluate the presence of BV, healthy and intermediate microbiota, by the criteria of Donders to determine the presence of AV and by the Marot-Leblond criteria to diagnose VVC. DNA extraction from vaginal samples and Polymerase Chain Reaction (PCR) analysis was performed to characterize the presence of Gardnerella spp., Mobiluncus mulieris, Escherichia coli, Enterococcus spp., and Lactobacillus spp. Finally, quantification of the lactobacilli was performed by quantitative real-time PCR (qPCR) for samples from women with normal vaginal microbiota and women with AV. Our results showed 52% of women with healthy microbiota, 7% with intermediate microbiota, and 41% with vaginal dysbiosis, comprising 27% with AV, 8% with BV and 4% with VVC and 2% with co-infections or co-dysbiosis. Additionally, a higher amount of lactobacilli were found in pregnant women when compared to non-pregnant women, while AV cases were characterized by a significant drop of Lactobacillus spp., more precisely, between 1E3 and 1E5 colony forming units (CFU)/ml. Finally, women with normal vaginal microbiota showed an average load of lactobacilli between 1E6 and 1E7 CFU/ml. This pilot study showed no statistically significant differences between pregnant and non-pregnant women, pointing to the possibility to use lactobacilli quantification for the prevention of future vaginal infections.



中文翻译:

通过 qPCR 对孕妇和非孕妇进行阴道微生物群评估和乳酸杆菌定量:一项试点研究。

妊娠结局和女性健康直接受到阴道微生物群的影响。该微生物群由不同比例的各种微生物组成的动态生态系统组成,在健康条件下可保护阴道上皮免受感染。然而,育龄妇女经常诊断出阴道感染病例,从而导致更严重的后果。因此,我们的主要目标是确定厄瓜多尔孕妇和非孕妇中细菌性阴道病 (BV)、需氧性阴道炎 (AV) 和外阴阴道念珠菌病 (VVC) 的患病率。2018 年 10 月至 2019 年 2 月期间,对 Carlos Andrade Marin 医院 (HCAM)、Isidro Ayora 妇产医院 (HGOIA) 和 Cipriana Dueñas 健康教学中心寻求初级保健的 217 名 13 至 40 岁女性进行了一项横断面研究阴道微生物群的经典表征是通过显微镜按照 Nugent 标准评估 BV、健康和中间微生物群的存在,按照 Donders 标准确定 AV 的存在,并按照 Marot-Leblond 标准诊断 VVC。从阴道样本中提取 DNA 并进行聚合酶链反应 (PCR) 分析,以表征是否存在加德纳菌种,Mobiluncus mulieris、大肠杆菌、肠球菌种,和乳酸菌种。最后,通过实时定量 PCR (qPCR) 对具有正常阴道微生物群的女性和患有 AV 的女性的样本进行乳酸杆菌定量。我们的结果显示,52% 的女性微生物群健康,7% 的女性微生物群处于中等水平,41% 的女性存在阴道菌群失调,其中 27% 患有 AV,8% 患有 BV,4% 患有 VVC,2% 患有合并感染或共同失调。此外,与非孕妇相比,孕妇体内的乳酸杆菌含量更高,而 AV 病例的特点是乳酸菌含量显着下降。乳酸菌更准确地说,在 1E3 和 1E5 菌落形成单位 (CFU)/ml 之间。最后,阴道微生物群正常的女性的平均乳酸菌负荷在 1E6 至 1E7 CFU/ml 之间。这项初步研究表明,孕妇和非孕妇之间没有统计学上的显着差异,这表明使用乳酸杆菌定量来预防未来阴道感染的可能性。

更新日期:2020-06-19
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