Front Microbiol . 2020 Mar 24;11:415. doi: 10.3389/fmicb.2020.00415. eCollection 2020.
Abstract
Infection and inflammation are well recognized causes of spontaneous preterm delivery (PTD) (<37 gestational weeks) and adverse infant outcomes. To date, there has been very little investigation into bacterial communities in amniotic fluid using next generation sequencing technology. In particular, it is important to characterize amniotic fluid bacterial profiles in complicated pregnancies as well as in asymptomatic women to identify predictive bacterial DNA signatures. Here, 1198 mid-trimester amniotic fluid samples from a cohort of Swedish women undergoing mid-trimester genetic amniocentesis were screened for bacterial DNA using qPCR protocols specifically designed to reduce the impacts of reagent contamination and human DNA mispriming. The majority of samples were devoid of detectable bacterial DNA; however, approximately a fifth of the cohort (19.9%) were 16S rRNA gene positive in duplicate screening. Among these, nine women had a spontaneous PTD. These nine women were matched with 18 healthy women with a delivery at term. We used PacBio SMRT technology, coupled with appropriate negative extraction and PCR controls, to sequence the full-length 16S rRNA gene in this subset of 27 women. The amniotic fluid samples contained low-abundance and low-diversity bacterial DNA profiles. Species typically associated with spontaneous PTD were absent. We were not able to identify any differences in the amniotic fluid bacterial DNA profiles of women with a subsequent spontaneous PTD compared to women who delivered at term. The findings suggest that, in a minor proportion of pregnancies, DNA from non-pathogenic bacteria may be present in the amniotic fluid far earlier than previously reported. Early detection of bacterial DNA in the amniotic fluid was, in this study, not associated with spontaneous PTD.
Keywords: 16S rRNA; amniotic fluid; bacteria; microbial invasion of amniotic cavity; preterm birth
研究背景
自发性早产(PTD)是指妊娠28-37周之间的分娩者,已有研究表明羊水中病原菌的定植是引发感染并导致PTD的原因,但对于羊水中的细菌性质、起源、定植时期和意义尚不明确,这些细菌种群在胎儿的皮肤和肠道定植,可能对胎儿的发育成熟和免疫系统产生重要影响。
图1. 队列流程图及受试人群选择
研究结果
1. 妊娠中期羊水中细菌DNA存在情况
研究未发现通常与子宫内感染和早产有关的细菌。支持了此类物种入侵可能发生在妊娠20周后的观点,本研究中确定的物种潜在定殖发生的时间更早,并且是非致病性的。结合“无菌子宫”理论,研究认为羊水中存在的细菌特征不一定代表真正的“微生物组”,仅仅反映了细菌从母亲转移到胎儿的过程,这些细菌在子宫内短暂存在但不发生定殖,从而导致了细菌低丰度的状态。
结论
研究结果表明,尽管大多数妊娠中期的羊水样本都是无菌的,但大约有五分之一样本中存在细菌DNA。妊娠中期羊水细菌DNA谱除具有低生物量和低多样性的特点外,还缺乏与PTD相关的物种,说明羊水中非病原性细菌存在可能远早于以往认为的时期。妊娠中期羊水样本中是否存在细菌DNA的发育和免疫学意义尚不清楚。