新型蚕丝产道微生态转移体的制备

  目的  优化新型蚕丝“产道微生态转移体（birth-canal microecology transporter, BMT）”制备参数，高效转移产道共生菌。  方法  纳入正常足月待产孕妇30例为研究对象。收集研究对象入院时产道微生物样本，作为对照组（NC，n=30）；其中18例最终选择剖宫产终止妊娠者纳入试验组（M），将其随机分成6个亚组（每组3例），进行“产道微生态转移”试验。于剖宫产术前准备时将新型蚕丝“BMT”以不同浸润度分别浸入不同渗透压的无菌液体中处理后，置入并贴附于孕妇阴道壁，保留1 h后取出封存。所有样本经DNA提取、扩增16S rDNA V3~V4区，应用 Illumina Hiseq2500进行测序。利用Mothur、QIIME、Lefse和Metastats等软件进行微生物多样性分析，采用Welch’s t检验及Anosim非参数检验等方法进行差异性分析。  结果  70%浸润度的蚕丝“BMT”在浸润瞬间可达溶液全覆盖，且保液性优，贴附效果佳，受试者无异物感，体验度好。新型“BMT”负载的微生物与正常妊娠末期阴道的微生物物种组成在属水平均以乳杆菌属为优势主导菌，且在0.45%无菌NaCl溶液、70%浸润度条件下，蚕丝“BMT”负载微生物的菌群构成、多样性信息与对照组最为接近，组间差异无统计学意义（P>0.05）。  结论  新型蚕丝“BMT”可高效转移产道共生菌，以0.45%低渗盐溶液、70%浸润度条件为转移体制备最优参数。

Preparation of a New Type of Silk Birth-canal Microecology Transporter

  Objective  To optimize the preparation parameters of the new silk birth-canal microecology transporter (BMT) for transferring the symbiotic bacteria of the birth canal efficiently.   Methods  Birth canal microbial samples of 30 full term pregnant women at admission were collected as the control group (NC, n=30). The experimental group included 18 pregnant women terminated by Cesarean section, who were divided into 6 sub-groups （M1-M6, n=3) to complete the transfer tests of the birth-canal microecology. The new silk BMT was processed in the sterile liquid of the different osmotic pressure with the different immersion depth, and was placed in the vagina of the pregnant women for 1 h before sealed. All extracted DNA specimens were amplified in the V3-V4 region of the 16S rDNA, and were sequenced by Illumina Hiseq2500. Microbial diversity analysis was performed by Mothur, QIIME, Lefse and Metastat. Welch’s t-test and Anosim nonparametric test were used to compare the difference between groups.  Results  The new silk BMT with 70% immersion depth could be fully covered by the solution, and had good solution preserving and adhesion. The subjects had no foreign body sensation with satisfied experience. Both of the microbes on the new BMT and the control group were lactobacillus as the dominant bacteria genus. The microbial diversity and bacteria constitution in the new BMT was similar to the control group in the condition of 0.45% NaCl solution and 70% immersion depth, and there was no significant difference between the two groups (P>0.05).   Conclusion  The new silk BMT can transfer the symbiotic microbes of the birth canal efficiently, and the optimal preparation parameters were 0.45% hypotonic saline solution and 70% immersion depth.