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采用 Sanger 测序法评估健康波兰女性妊娠后三个时期阴道和直肠乳杆菌属菌群的动态变化。

The dynamics of vaginal and rectal Lactobacillus spp. flora in subsequent trimesters of pregnancy in healthy Polish women, assessed using the Sanger sequencing method.

机构信息

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, Czysta 18, 31-121, Krakow, Poland.

Clinical Department of Gynecological Endocrinology and Gynecology, Jagiellonian University Medical College, Kopernika 23, 31-501, Krakow, Poland.

出版信息

BMC Pregnancy Childbirth. 2018 Aug 29;18(1):350. doi: 10.1186/s12884-018-1987-7.

Abstract

BACKGROUND

Lactobacilli play an important role in maintaining vaginal health and protection against bacterial infections in the genital tract. The aim of this study is to show the dynamics of changes of the vaginal and rectal Lactobacillus flora during pregnancy by using the Sanger sequencing method.

METHOD

The study included 31 healthy pregnant women without clinical signs of genitourinary infections. The material was taken in the three trimesters of pregnancy by vaginal and rectal swabs and grown on the MRS agar quantitatively to estimate the number of Lactobacillus spp. [CFU/ml]. Afterwards, 3 to 8 morphologically different lactobacilli colonies were taken for identification. Bacterial species identification was performed by 16 s rDNA sequence fragment analyses using the Sanger method.

RESULTS

Among the patients tested, the most common species colonizing the vagina in the first trimester were: L. crispatus 29%, L. gasseri 19.4% and L. rhamnosus 16.1%, in the second trimester: L. crispatus 51.6%, L. gasseri 25.8%, L. rhamnosus 19.4% and L. amylovorus 16.1%, and in the third trimester the most common Lactobacillus species were: L. crispatus 25.8%, L. gasseri 25.8% and L. johnsonii 19.4%. In rectal species, the number decreased in the second and third trimesters in comparison to the first trimester (p = 0.003). An analysis of rectal dynamics showed that in the first trimester, the most common species were: L. johnsonii 19.4%, and L. plantarum 9.7%, in the second trimester: L. crispatus 9.7% and L. mucosae 6.5%, and in the third trimester: L. casei 9.7% and L. rhamnosus 9.7%. Individual dynamics of the Lactobacillus species composition showed variability, characterized by continuous, intermittent, or periodic colonization. The patients examined were mostly colonized by three Lactobacillus species in vagina (32.3%), whereas for the rectum, one Lactobacillus species during the whole pregnancy duration was common (32.3%).

CONCLUSION

This study showed that in the examined group of healthy, pregnant Polish women, the vaginal Lactobacillus flora, both qualitative and quantitative, was stable during the three subsequent trimesters. In contrast, the number of rectal Lactobacillus species dramatically decreased after the first trimester.

摘要

背景

乳酸菌在维持阴道健康和预防生殖道细菌感染方面发挥着重要作用。本研究旨在通过 Sanger 测序法展示怀孕期间阴道和直肠乳酸菌菌群变化的动态。

方法

该研究纳入了 31 名无泌尿生殖道感染临床症状的健康孕妇。通过阴道和直肠拭子在妊娠的三个阶段采集标本,并在 MRS 琼脂上定量生长以估计乳酸菌 spp. 的数量[CFU/ml]。之后,从 3 到 8 个形态不同的乳酸菌菌落中取出用于鉴定。细菌种属鉴定采用 Sanger 法对 16s rDNA 序列片段进行分析。

结果

在所测试的患者中,在第一个三个月中最常见的定植阴道的菌种为:L. crispatus(29%)、L. gasseri(19.4%)和 L. rhamnosus(16.1%);在第二个三个月中为:L. crispatus(51.6%)、L. gasseri(25.8%)、L. rhamnosus(19.4%)和 L. amylovorus(16.1%);在第三个三个月中最常见的乳酸菌种属为:L. crispatus(25.8%)、L. gasseri(25.8%)和 L. johnsonii(19.4%)。在直肠中,与第一个三个月相比,第二和第三个三个月的数量减少(p=0.003)。对直肠动态的分析表明,在第一个三个月中,最常见的菌种为:L. johnsonii(19.4%)和 L. plantarum(9.7%);在第二个三个月中为:L. crispatus(9.7%)和 L. mucosae(6.5%);在第三个三个月中为:L. casei(9.7%)和 L. rhamnosus(9.7%)。乳酸菌种属组成的个体动态表现出变异性,其特征为连续、间歇或周期性定植。所检查的患者阴道中大多定植有三种乳酸菌(32.3%),而直肠中整个妊娠期间常见一种乳酸菌(32.3%)。

结论

本研究表明,在检查的波兰健康孕妇组中,阴道乳酸菌菌群在随后的三个三个月中无论是定性还是定量均保持稳定。相比之下,直肠乳酸菌的数量在第一个三个月后急剧减少。

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