Objective
To investigate the results of high-throughput sequencing of placenta and membrane flora abundance in premature rupture of membranes (PROM)and normal pregnant women,and to analyze the differences.
Methods
Fifteen pregnant women who gave birth in the Department of Obstetrics,the First Affiliated Hospital of Kunming Medical University from January 2019 to February 2020 were selected as study subjects.According to whether PROM occurred or not and their gestational age at the time of PROM occurrence,they were included into preterm PROM(PPROM)group(n=5,PPROM pregnant women),term PROM (TPROM)group(n=6,TPROM pregnant women)and control group(n=4,pregnant women without PROM).Immediately after placenta delivery,one sample of tissue from fetal face and maternal face placenta and fetal membrane was collected from pregnant women in three groups.The flora DNA in placenta and fetal membrane tissues was extracted,and the high variable region V3-V4 of 16S ribosomal RNA(r RNA)gene was sequenced with high throughput,and the flora structure of phylum and genus in the placenta and fetal membrane tissue samples of pregnant women in three groups was analyzed.Metastats software was used to compare the relative abundance of bacteria in placenta and fetal membranes tissues in PPROM group and control group,TPROM group and control group,PPROM group and TPROM group by parameter T test.There were no significant differences in the general clinical data such as age and weight of pregnant women during delivery among three groups (P >0.05).The procedure followed in this study met the requirements of the WorldMedicalAssociationDeclaration ofHelsinki revised in 2013.
Results
①The difference in gestational age of pregnant women among three groups was statistically significant(F=18.65,P<0.001).②At the phylum level,there were 7 phyla with relative abundance>0.1%in the placental and fetal membrane tissue samples of pregnant women in three groups:Firmicutes,Bacteroidetes,Spirochaetes,Proteobacteria,Actinobacteria,Cyanobacteria,and Fusobacteria.③At the genus level,there were 6 species of bacteria with relative abundance>0.1%in the placental and fetal membrane tissue samples of pregnant women in three groups:uncultured bacterium,Lactobacillus,Clostridium,Streptococcus,Catenibacterium,Treponema.④The difference analysis of genus level bacterial flora in placenta and fetal membrane tissue samples of pregnant women in three groups showed that the relative abundance of Clostridium,Streptococcus,unclassified bacteria and Treponema in placenta and fetal membrane tissue samples of pregnant women in PPROM and TPROM groups were significantly higher than those in control group,while the relative abundance of Streptobacillus in PPROM and TPROM groups were significantly lower than that in control group,and all the differences were statistically significant (P<0.05).
Conclusions
The placenta and fetal membranes of normal pregnant women and PPROM and TPROM pregnant women are not sterile,and the distribution of placenta and fetal membrane flora among them is diverse.Compared with normal pregnant women,the diversity of placenta and fetal membrane flora of PROM pregnant women is consistent,but the abundance of some flora is obviously different.