卵泡液和胚胎培养废弃培养液中 cfDNA 的完整性与体外受精患者胚胎等级有关。
The integrity of cfDNA in follicular fluid and spent medium from embryo culture is associated with embryo grade in patients undergoing in vitro fertilization.
机构信息
School of Medicine, Southeast University, Nanjing, China.
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China.
出版信息
J Assist Reprod Genet. 2021 Dec;38(12):3113-3124. doi: 10.1007/s10815-021-02357-0. Epub 2021 Nov 24.
PURPOSE
This study was conducted to verify if the cfDNA integrity (cfDI) in follicular fluid and subsequent spent embryo medium (SEM) could serve as potential non-invasive biomarker for high-grade embryo selection during IVF/ICSI.
METHODS
Thirty-two follicular fluids, 32 subsequent corresponding cleavage embryo SEM, and 23 subsequent blastocyst SEM were collected from 11 patients undergoing IVF/ICSI. CfDI was measured by ALU gene amplicons with different sizes by qPCR, as the ratio of long to short fragments.
RESULTS
CfDI in follicular fluid corresponding to subsequent high-grade cleavage embryos and blastocysts was significantly lower than that related to low-grade embryos (p = 0.018). Conversely, cfDI in SEM was significantly and positively correlated with high-grade embryos at both stages (p = 0.009). ROC curves of the analysis of cfDI in follicular fluid showed great potential in predicting subsequent embryogenesis and embryo grade (AUC > 0.927). Regardless of the cleavage embryo grade by morphology, cfDI in day 3 SEM could predict if the cleavage embryo could develop to a high-grade blastocyst (AUC = 0.820). A concordant shift pattern of cfDI from follicular fluid to subsequent day 3 SEM and day 5 SEM was found in 81.82% participants featured by various clinical characteristics.
CONCLUSION
CfDI in follicular fluid and SEM was significantly correlated with embryogenesis and embryo grade and could serve as a potential non-invasive biomarker in high-grade embryo selection. Direct qPCR was proved as a labor-saving and sensitive method for the analysis of cfDI in low volume of SEM.
目的
本研究旨在验证卵泡液和随后的胚胎培养液中的 cfDNA 完整性（cfDI）是否可作为 IVF/ICSI 中选择优质胚胎的潜在非侵入性生物标志物。
方法
从 11 名接受 IVF/ICSI 的患者中收集了 32 个卵泡液、32 个随后的卵裂胚胎 SEM 和 23 个随后的囊胚 SEM。通过 qPCR 用不同大小的 ALU 基因扩增子测量 cfDI，作为长片段与短片段的比值。
结果
与低质量胚胎相比，卵泡液中与后续高质量卵裂胚胎和囊胚相关的 cfDI 显著降低（p = 0.018）。相反，在两个阶段，SEM 中的 cfDI 与高质量胚胎呈显著正相关（p = 0.009）。卵泡液 cfDI 分析的 ROC 曲线显示出在预测后续胚胎发生和胚胎等级方面具有很大的潜力（AUC > 0.927）。无论形态学上的卵裂胚胎等级如何，第 3 天 SEM 中的 cfDI 都可以预测卵裂胚胎是否可以发育成高质量的囊胚（AUC = 0.820）。在具有各种临床特征的 81.82%的参与者中，发现了 cfDI 从卵泡液到随后的第 3 天 SEM 和第 5 天 SEM 的一致转移模式。
结论
卵泡液和 SEM 中的 cfDI 与胚胎发生和胚胎等级显著相关，可作为选择优质胚胎的潜在非侵入性生物标志物。直接 qPCR 被证明是一种节省劳力和敏感的方法，用于分析低体积 SEM 中的 cfDI。
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