Endometrial microbiome at the time of embryo transfer: next-generation sequencing of the 16S ribosomal subunit
Endometrial microbiome at the time of embryo transfer: next-generation sequencing of the 16S ribosomal subunit
复制标题
DOI：
10.1007/s10815-015-0614-z
复制
发表时间：
2016-01-01
影响因子：
3.1
通讯作者：
Scott, R. T.
中科院分区：
文献类型：
--
作者：
Franasiak, J. M.;Werner, M. D.;Scott, R. T.
随着强大的分子工具的应用，人类微生物组的表征变得更加精确，这些工具利用独特的16S核糖体亚基的高变区来大大提高灵敏度。下生殖道的微生物组可以预测产科结局，而上生殖道的特征仍然很差。在此，单胚胎移植（SET）时子宫内膜微生物组的特征是生殖结果。接受整倍体SET的妊娠患者被纳入分析。胚胎移植后，按照常规进行，将移植导管的最远端5 mm部分无菌放置在无DNA PCR管中。对细菌特异性16S核糖体基因进行了下一代测序，允许对微生物进行属和种的调用。对来自33名患者和2名大肠杆菌对照的35份样品进行了分类学分配。在33名患者中，18名正在怀孕，15名没有怀孕。在患者样本中总共存在278个不同的属调用。正在进行妊娠的患者与没有正在进行妊娠的患者在转移时的微生物组的特征在于按总和分数计算的最高属。乳酸菌是这两种结果的首选菌种。这里提供的数据表明，胚胎移植时的微生物组可以在不改变标准临床实践的情况下成功地进行表征。这种新的方法，无论是在标本收集和分析，是朝着确定生理从病理生理微生物群的目标的第一步。进一步的研究将有助于阐明胚胎移植时微生物组的差异是否对妊娠结果产生可靠的影响。
Characterization of the human microbiome has become more precise with the application of powerful molecular tools utilizing the unique 16S ribosomal subunit's hypervariable regions to greatly increase sensitivity. The microbiome of the lower genital tract can prognosticate obstetrical outcome while the upper reproductive tract remains poorly characterized. Here, the endometrial microbiome at the time of single embryo transfer (SET) is characterized by reproductive outcome.Consecutive patients undergoing euploid, SET was included in the analysis. After embryo transfer, performed as per routine, the most distal 5-mm portion of the transfer catheter was sterilely placed in a DNA free PCR tube. Next-generation sequencing of the bacteria specific 16S ribosome gene was performed, allowing genus and species calls for microorganisms.Taxonomy assignments were made on 35 samples from 33 patients and 2 Escherichia coli controls. Of the 33 patients, 18 had ongoing pregnancies and 15 did not. There were a total of 278 different genus calls present across patient samples. The microbiome at time of transfer for those patients with ongoing pregnancy vs. those without ongoing pregnancy was characterized by top genera by sum fraction. Lactobacillus was the top species call for both outcomes.The data presented here show the microbiome at the time of embryo transfer can successfully be characterized without altering standard clinical practice. This novel approach, both in specimen collection and analysis, is the first step toward the goal of determining physiologic from pathophysiologic microbiota. Further studies will help delineate if differences in the microbiome at the time of embryo transfer have a reliable impact on pregnancy outcome.