School of Medicine, Southeast University, Nanjing, China.
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China.
J Assist Reprod Genet. 2021 Dec;38(12):3113-3124. doi: 10.1007/s10815-021-02357-0. Epub 2021 Nov 24.
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.
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.
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.
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.
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。
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。
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。
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。
