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Isolation of Gardnerella vaginalis from BV patients and healthy women : analysis of virulence through adherence, biofilm formation and cytotoxicity assays

机译:从BV患者和健康女性中分离阴道加德纳菌:通过黏附,生物膜形成和细胞毒性测定分析毒力

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摘要

Bacterial vaginosis (BV) is one of the most common gynaecological disorder affecting women in the reproductive age. Microbiological analysis of BV has shown Gardnerella vaginalis to be the most frequent organism in BV. However, G. vaginalis colonization do not always lead to BV. This raised the question whether there are pathogenic and commensal lineages within this species. In an effort to understand the differences between G. vaginalis strains, we performed in vitro assays to compare virulence properties of recently isolated 14 G. vaginalis strains from Portuguese women with and without BV. G. vaginalis strains were characterised for their initial adhesion ability to a monolayer of HeLa cells by incubating the bacteria with this monolayer and quantifying the adhesion by staining with DAPI and fluorescence microscopy. These assays revealed that the BV isolates of G. vaginalis had a stronger initial adhesion capability than non-BV isolates. The biofilm-forming capacity was then assessed by allowing each of the strains to form biofilms under anaerobic conditions for 48 hours and using different growth media. It was possible to observe that BV isolates tend to growth preferentially as biofilms while non-BV isolates had a lower intrinsic tendency towards biofilm formation. In addition, BV isolates of G. vaginalis displayed robust cytotoxicity in the epithelial cells after 3 hours in the contact with a monolayer of HeLa cells. Thus, this study outlines two distinct variants of G. vaginalis, one apparently commensal and one pathogenic, and presents evidence for disparate virulence potentials.
机译:细菌性阴道病(BV)是影响生育年龄妇女的最常见的妇科疾病之一。 BV的微生物学分析表明阴道加德纳菌是BV中最常见的生物。但是,阴道加德纳菌定植并不总是导致BV。这就提出了一个问题,即该物种内是否有致病性和共生谱系。为了理解阴道G.G.菌株之间的差异,我们进行了体外测定,以比较最近分离出的来自葡萄牙妇女有和没有BV的14 G.阴道G.菌株的毒力特性。通过将细菌与单层HeLa一起孵育,并用DAPI染色和荧光显微镜法定量其粘附力,来表征其对HeLa细胞单层的初始粘附能力。这些测定法揭示了阴道假单胞菌的BV分离株比非BV分离株具有更强的初始粘附能力。然后通过使每个菌株在厌氧条件下48小时并使用不同的生长培养基形成生物膜来评估生物膜形成能力。可以观察到BV分离株倾向于优先生长为生物膜,而非BV分离株具有较低的生物膜形成内在趋势。此外,与单层HeLa细胞接触3小时后,阴道加德纳氏菌的BV分离株在上皮细胞中显示出强大的细胞毒性。因此,这项研究概述了阴道丙种球菌的两种不同变体,一种明显是共生的,一种是致病性的,并提供了不同毒力潜能的证据。

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