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证据表明,羊膜内感染通常是上行性感染的结果——一项分子微生物学研究。

Evidence that intra-amniotic infections are often the result of an ascending invasion - a molecular microbiological study.

机构信息

Perinatology Research Branch, Division of Obstetrics and Maternal-Fetal Medicine, Division of Intramural Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, MD and Detroit, MI, USA.

Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, MI, USA.

出版信息

J Perinat Med. 2019 Nov 26;47(9):915-931. doi: 10.1515/jpm-2019-0297.

Abstract

Background Microbial invasion of the amniotic cavity resulting in intra-amniotic infection is associated with obstetrical complications such as preterm labor with intact or ruptured membranes, cervical insufficiency, as well as clinical and histological chorioamnionitis. The most widely accepted pathway for intra-amniotic infection is the ascension of microorganisms from the lower genital tract. However, hematogenous dissemination of microorganisms from the oral cavity or intestine, retrograde seeding from the peritoneal cavity through the fallopian tubes, and introduction through invasive medical procedures have also been suggested as potential pathways for intra-amniotic infection. The primary reason that an ascending pathway is viewed as most common is that the microorganisms most often detected in the amniotic fluid are those that are typical inhabitants of the vagina. However, thus far, no studies have shown that microorganisms in the amniotic cavity are simultaneously present in the vagina of the woman from which they were isolated. The objective of the study was to determine the frequency with which microorganisms isolated from women with intra-amniotic infection are also present in the lower genital tract. Methods This was a cross-sectional study of women with intra-amniotic infection with intact membranes. Intra-amniotic infection was defined as a positive culture and elevated concentrations of interleukin-6 (IL-6) (>2.6 ng/mL) in amniotic fluid and/or acute histologic chorioamnionitis and funisitis. Microorganisms isolated from bacterial cultures of amniotic fluid were taxonomically identified through matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) and 16S ribosomal RNA (rRNA) gene sequencing. Vaginal swabs were obtained at the time of amniocentesis for the identification of microorganisms in the lower genital tract. The overall bacterial profiles of amniotic fluids and vaginal swabs were characterized through 16S rRNA gene sequencing. The bacterial profiles of vaginal swabs were interrogated for the presence of bacteria cultured from amniotic fluid and for the presence of prominent (>1% average relative abundance) operational taxonomic units (OTUs) within the overall 16S rRNA gene bacterial profiles of amniotic fluid. Results (1) A total of 75% (6/8) of women had bacteria cultured from their amniotic fluid that are typical residents of the vaginal ecosystem. (2) A total of 62.5% (5/8) of women with bacteria cultured from their amniotic fluid also had these bacteria present in their vagina. (3) The microorganisms cultured from amniotic fluid and also detected in the vagina were Ureaplasma urealyticum, Escherichia coli, and Streptococcus agalactiae. (4) 16S rRNA gene sequencing revealed that the amniotic fluid of women with intra-amniotic infection had bacterial profiles dominated by Sneathia, Ureaplasma, Prevotella, Lactobacillus, Escherichia, Gardnerella, Peptostreptococcus, Peptoniphilus, and Streptococcus, many of which had not been cultured from the amniotic fluid samples. (5) Seventy percent (7/10) of the prominent (>1% average relative abundance) OTUs found in amniotic fluid were also prominent in the vagina. Conclusion The majority of women with intra-amniotic infection had bacteria cultured from their amniotic fluid that were typical vaginal commensals, and these bacteria were detected within the vagina at the time of amniocentesis. Molecular microbiological interrogation of amniotic fluid from women with intra-amniotic infection revealed that the bacterial profiles of amniotic fluid were largely consistent with those of the vagina. These findings indicate that ascension from the lower genital tract is the primary pathway for intra-amniotic infection.

摘要

背景

微生物侵入羊膜腔导致羊膜腔内感染与早产、胎膜完整或破裂、宫颈功能不全以及临床和组织学绒毛膜羊膜炎等产科并发症有关。羊膜腔内感染最广泛接受的途径是微生物从下生殖道上行。然而,微生物也可能通过血源性播散从口腔或肠道进入,通过输卵管逆行种植从腹腔进入,以及通过侵入性医疗程序引入。之所以认为上行途径最常见,主要原因是在羊水中最常检测到的微生物是阴道的典型居民。然而,迄今为止,尚无研究表明羊膜腔内的微生物与从其分离的女性阴道中的微生物同时存在。本研究的目的是确定从羊膜腔内感染的女性中分离出的微生物在生殖道下段同时存在的频率。

方法

这是一项对胎膜完整的羊膜腔内感染女性的横断面研究。羊膜腔内感染定义为羊水细菌培养阳性且白细胞介素-6(IL-6)浓度升高(>2.6ng/mL),以及急性组织学绒毛膜羊膜炎和脐带炎。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)和 16S 核糖体 RNA(rRNA)基因测序对羊水细菌培养物中分离出的微生物进行分类鉴定。在羊膜穿刺术时获取阴道拭子,以鉴定生殖道下段的微生物。通过 16S rRNA 基因测序对羊水和阴道拭子的整体细菌谱进行特征描述。对阴道拭子的细菌谱进行分析,以确定是否存在从羊水培养出的细菌,以及是否存在羊水总体 16S rRNA 基因细菌谱中突出的(平均相对丰度>1%)分类单元(OTU)。

结果

(1)共有 6/8(75%)名女性的羊水培养出了阴道生态系统的典型居民细菌。(2)共有 5/8(62.5%)名羊水培养出细菌的女性阴道内也存在这些细菌。(3)从羊水和阴道中培养出的微生物为 Ureaplasma urealyticum、Escherichia coli 和 Streptococcus agalactiae。(4)16S rRNA 基因测序显示,羊膜腔内感染女性的羊水细菌谱以 Sneathia、Ureaplasma、Prevotella、Lactobacillus、Escherichia、Gardnerella、Peptostreptococcus、Peptoniphilus 和 Streptococcus 为主,其中许多细菌未从羊水样本中培养出来。(5)在羊水样本中发现的 70%(7/10)个突出的(平均相对丰度>1%)OTU 也在阴道中突出。

结论

大多数羊膜腔内感染的女性羊水培养出的细菌都是阴道的典型共生菌,在羊膜穿刺术时这些细菌存在于阴道内。对羊膜腔内感染女性的羊水进行分子微生物学检测发现,羊水的细菌谱与阴道的细菌谱基本一致。这些发现表明,从生殖道下段上行是羊膜腔内感染的主要途径。

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