Reproductive Medicine Unit, S.I.S.Me.R., Società Italiana Studi Medicina della Riproduzione, Bologna, Italy.
Hum Reprod. 2023 Jul 5;38(7):1268-1276. doi: 10.1093/humrep/dead088.
Is the presence of DNA in the blastocoel fluid (BF) of expanded blastocysts, assessed by whole genome amplification (WGA), associated with the clinical outcome at the first transfer?
At the first transfer, blastocysts with negative BF-WGA have more chance to implant and to develop to term than those with positive BF-WGA results, both in preimplantation genetic testing for aneuploidies (PGT-A) cycles (where only euploid blastocysts resulting from the chromosomal analysis of trophectoderm (TE) biopsies were transferred) and in IVF/ICSI conventional cycles.
Retrospective studies conducted in patients undergoing PGT-A have shown that the incidence of negative BF-WGA was significantly higher in TE-euploid blastocysts than in TE-aneuploid blastocysts. In addition, after the transfer of TE-euploid blastocysts, the ongoing clinical pregnancy rate was significantly higher in the group with negative BF-WGA compared with those with positive BF-WGA.
STUDY DESIGN, SIZE, DURATION: A prospective cohort study including 102 consecutive PGT-A patients (Group 1) and 88 consecutive conventional IVF/ICSI patients (Group 2), was conducted between January 2019 and December 2021.
PARTICIPANTS/MATERIALS, SETTING, METHODS: In both groups, BFs were collected from expanded blastocysts of high grade and processed for WGA. DNA amplification was evaluated by agarose gel electrophoresis for the presence (positive BF-WGA) or absence (negative BF-WGA) of a band. Directly after the BF retrieval, blastocysts from Group 1 underwent TE biopsy and vitrification. In Group 2, blastocysts were vitrified immediately after BF collection. In Group 1, only euploid blastocysts were considered for transfer according to the results of TE biopsies. In both groups, the selection of the blastocyst to be transferred was based on BF-WGA results giving priority, if available, to those with negative amplification. The primary outcome investigated was the live birth rate (LBR) at the first transfer. The main variable under investigation was the negative BF-WGA and results were corrected for confounders (maternal and paternal age, number of retrieved oocytes, male factor) by multiple logistic regression analysis.
In Group 1, 60 patients transferred negative BF-WGA blastocysts and 42 positive BF-WGA blastocysts, and the LBR at the first transfer was 53.3% and 26.2%, respectively (P = 0.0081). After testing for selected confounders in a multiple logistic analysis, the transfer of blastocysts with negative BF-WGA resulted in an odds ratio of (OR) 3.52 (95% CI: 1.48-8.88, P = 0.0057) compared to transfer of positive BF-WGA blastocysts. In Group 2, at the first transfer 30 deliveries resulted from blastocysts with negative BF-WGA (48.4%) and three from the transfer of positive BF-WGA blastocysts in 26 patients (11.5%; P = 0.0014). Multiple logistic analysis indicated that the transfer of blastocysts with negative BF-WGA resulted in an OR 6.89 (95% CI: 1.98-32.95, P = 0.0056) compared to transfer of positive BF-WGA blastocysts. The LBR per transfer and the cumulative LBR per patient showed the same trend.
LIMITATIONS, REASONS FOR CAUTION: The study was performed in a single center.
The data from this study highlight the heterogeneity of blastocysts of similar morphology, even in those classified as euploid by TE analysis. Failure to detect DNA in BFs after WGA is associated with a significantly higher LBR at the first embryo transfer as well as per transfer and per patient. The processing of the BF by WGA is an easy and cost-effective tool that could become a valuable option to offer patients the highest chances of term pregnancy in the shortest time possible.
STUDY FUNDING/COMPETING INTEREST(S): The study received no funding from external sources. There are no conflicts of interest to declare.
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通过全基因组扩增（WGA）评估扩张囊胚腔液（BF）中是否存在 DNA 是否与首次移植的临床结局相关？
在首次移植中，与 BF-WGA 阳性结果相比，BF-WGA 阴性的囊胚具有更高的着床和发育至足月的机会，无论是在植入前遗传学检测非整倍体（PGT-A）周期（仅转移经滋养外胚层（TE）活检分析为整倍体的囊胚）还是在常规 IVF/ICSI 周期中。
在接受 PGT-A 的患者中进行的回顾性研究表明，在 TE 整倍体囊胚中，BF-WGA 阴性的发生率明显高于 TE 非整倍体囊胚。此外，在转移 TE 整倍体囊胚后，BF-WGA 阴性组的持续临床妊娠率明显高于 BF-WGA 阳性组。
研究设计、规模、持续时间：这项前瞻性队列研究纳入了 2019 年 1 月至 2021 年 12 月期间的 102 名连续接受 PGT-A 的患者（第 1 组）和 88 名连续接受常规 IVF/ICSI 的患者（第 2 组）。
参与者/材料、设置、方法：在两组中，均从高等级扩张囊胚中收集 BF 并进行 WGA。通过琼脂糖凝胶电泳评估 DNA 扩增情况，以确定是否存在（阳性 BF-WGA）或不存在（阴性 BF-WGA）带。在第 1 组中，直接在 BF 回收后进行 TE 活检和玻璃化处理。在第 2 组中，囊胚在收集 BF 后立即进行玻璃化处理。在第 1 组中，仅根据 TE 活检结果选择可转移的整倍体囊胚。在两组中，选择要转移的囊胚均基于 BF-WGA 结果，如果有，优先选择扩增阴性的囊胚。主要研究结果为首次移植的活产率（LBR）。主要研究变量为阴性 BF-WGA 结果，并通过多变量逻辑回归分析对母亲和父亲的年龄、获得的卵子数量、男性因素等混杂因素进行校正。
在第 1 组中，60 名患者转移了阴性 BF-WGA 囊胚，42 名患者转移了阳性 BF-WGA 囊胚，首次移植的 LBR 分别为 53.3%和 26.2%（P=0.0081）。在多变量逻辑分析中测试了选定的混杂因素后，与转移阳性 BF-WGA 囊胚相比，转移阴性 BF-WGA 囊胚的优势比（OR）为 3.52（95%CI：1.48-8.88，P=0.0057）。在第 2 组中，在首次移植时，30 次分娩来自阴性 BF-WGA 囊胚（48.4%），而 26 名患者中有 3 次分娩来自阳性 BF-WGA 囊胚（11.5%；P=0.0014）。多变量逻辑分析表明，与转移阳性 BF-WGA 囊胚相比，转移阴性 BF-WGA 囊胚的 OR 为 6.89（95%CI：1.98-32.95，P=0.0056）。每次转移的 LBR 和每位患者的累积 LBR 均显示出相同的趋势。
局限性、谨慎的原因：该研究仅在一个中心进行。
这项研究的数据强调了即使在 TE 分析中被归类为整倍体的囊胚中，囊胚的异质性。在 WGA 后未检测到 BF 中的 DNA 与首次胚胎移植以及每次移植和每位患者的较高 LBR 显著相关。BF 通过 WGA 处理是一种简单且具有成本效益的工具，可以为患者提供在最短时间内获得足月妊娠的最佳机会。
研究资金/利益冲突：该研究未获得外部来源的资金。没有利益冲突需要申报。
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