Non-invasive preimplantation genetic testing for aneuploidy using cell-free DNA in blastocyst culture medium.

IF 2.7 3区医学 Q2 GENETICS & HEREDITY

Journal of Assisted Reproduction and Genetics Pub Date : 2025-08-01 Epub Date: 2025-05-21 DOI:10.1007/s10815-025-03510-9

Ha T T Nguyen, Tam T M Luu, Linh T Đo, Tri C Nguyen, Diem T N Nguyen, Trang T M Ho, Hoa Giang, Thuy T H Dao, Bao G Huynh, Tuong M Ho, Lan N Vuong

{"title":"Non-invasive preimplantation genetic testing for aneuploidy using cell-free DNA in blastocyst culture medium.","authors":"Ha T T Nguyen, Tam T M Luu, Linh T Đo, Tri C Nguyen, Diem T N Nguyen, Trang T M Ho, Hoa Giang, Thuy T H Dao, Bao G Huynh, Tuong M Ho, Lan N Vuong","doi":"10.1007/s10815-025-03510-9","DOIUrl":null,"url":null,"abstract":"Purpose: This study evaluated the performance of preimplantation genetic testing for aneuploidies (PGT-A) using cell-free DNA (cfDNA) from spent culture media (SCM) of blastocyst embryos (non-invasive PGT-A; NiPGT-A) compared with conventional trophectoderm (TE) biopsy samples.Methods: This prospective study was conducted at IVFMD, My Duc Hospital, Vietnam, from August to December 2020, and included patients with an indication for PGT-A. The culture medium was replaced on day 3, and SCM from day 3 to the day of TE biopsy (days 5 or 6) of all biopsied blastocysts was tested using next-generation sequencing. The total concordance rate, sensitivity, and specificity of NiPGT-A versus PGT-A for detecting aneuploid embryos were calculated. Outcomes after single blastocyst transfer are also reported.Results: Forty-four couples participated; 100 paired TE PGT-A biopsies and SCM samples were evaluated. The whole-genome amplification success rate for SCM was 82%; 77 samples had clear NGS results and were further evaluated. The total concordance rate between NiPGT-A and PGT-A was 63.6%. For detecting aneuploidy, NiPGT-A had a sensitivity of 57.1%, specificity of 67.3%, positive predictive value of 50.0%, and negative predictive value of 73.3%. Of the 35 single euploidy embryo transfers, 8 had no NiPGT-A results, 21 were classified as NiPGT-A euploid, and 6 were classified as NiPGT-A aneuploid; the live birth rate was 51.4% (18/35). Four of the 6 NiPGT-A aneuploid blastocysts resulted in live births.Conclusions: cfDNA in SCM has the potential for NiPGT-A. However, the NiPGT-A process is unreliable enough to replace traditional PGT-A using TE biopsy.","PeriodicalId":15246,"journal":{"name":"Journal of Assisted Reproduction and Genetics","volume":" ","pages":"2587-2595"},"PeriodicalIF":2.7000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12423013/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Assisted Reproduction and Genetics","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s10815-025-03510-9","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/5/21 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}

引用次数: 0

Abstract

Purpose: This study evaluated the performance of preimplantation genetic testing for aneuploidies (PGT-A) using cell-free DNA (cfDNA) from spent culture media (SCM) of blastocyst embryos (non-invasive PGT-A; NiPGT-A) compared with conventional trophectoderm (TE) biopsy samples.

Methods: This prospective study was conducted at IVFMD, My Duc Hospital, Vietnam, from August to December 2020, and included patients with an indication for PGT-A. The culture medium was replaced on day 3, and SCM from day 3 to the day of TE biopsy (days 5 or 6) of all biopsied blastocysts was tested using next-generation sequencing. The total concordance rate, sensitivity, and specificity of NiPGT-A versus PGT-A for detecting aneuploid embryos were calculated. Outcomes after single blastocyst transfer are also reported.

Results: Forty-four couples participated; 100 paired TE PGT-A biopsies and SCM samples were evaluated. The whole-genome amplification success rate for SCM was 82%; 77 samples had clear NGS results and were further evaluated. The total concordance rate between NiPGT-A and PGT-A was 63.6%. For detecting aneuploidy, NiPGT-A had a sensitivity of 57.1%, specificity of 67.3%, positive predictive value of 50.0%, and negative predictive value of 73.3%. Of the 35 single euploidy embryo transfers, 8 had no NiPGT-A results, 21 were classified as NiPGT-A euploid, and 6 were classified as NiPGT-A aneuploid; the live birth rate was 51.4% (18/35). Four of the 6 NiPGT-A aneuploid blastocysts resulted in live births.

Conclusions: cfDNA in SCM has the potential for NiPGT-A. However, the NiPGT-A process is unreliable enough to replace traditional PGT-A using TE biopsy.

查看原文本刊更多论文

在胚泡培养基中使用无细胞DNA进行非整倍体植入前基因检测。

目的：本研究利用胚泡胚废培养基（SCM）中的无细胞DNA （cfDNA）对非整倍体（PGT-A）进行着床前遗传学检测(non-invasive PGT-A；NiPGT-A)与传统的滋养外胚层（TE）活检样本进行比较。方法：这项前瞻性研究于2020年8月至12月在越南myduc医院IVFMD进行，纳入了有PGT-A指征的患者。第3天更换培养基，使用下一代测序技术检测从第3天到TE活检当天（第5天或第6天）所有活检囊胚的SCM。计算NiPGT-A与PGT-A检测非整倍体胚胎的总一致性率、敏感性和特异性。单囊胚移植后的结果也有报道。结果：44对夫妇参与；对100例配对TE PGT-A活检和SCM样本进行评估。SCM全基因组扩增成功率为82%；77个样本有明确的NGS结果，并进行进一步评价。NiPGT-A与PGT-A的总符合率为63.6%。NiPGT-A检测非整倍体的敏感性为57.1%，特异性为67.3%，阳性预测值为50.0%，阴性预测值为73.3%。在35例单整倍体胚胎移植中，8例无NiPGT-A结果，21例为NiPGT-A整倍体，6例为NiPGT-A非整倍体；活产率为51.4%（18/35）。6个NiPGT-A非整倍体囊胚中有4个导致活产。结论：SCM患者的cfDNA具有诱发NiPGT-A的潜力。然而，NiPGT-A过程是不可靠的，足以取代传统的PGT-A使用TE活检。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of Assisted Reproduction and Genetics 医学-妇产科学

CiteScore

5.70

自引率

9.70%

发文量

286

审稿时长

1 months

期刊介绍： The Journal of Assisted Reproduction and Genetics publishes cellular, molecular, genetic, and epigenetic discoveries advancing our understanding of the biology and underlying mechanisms from gametogenesis to offspring health. Special emphasis is placed on the practice and evolution of assisted reproduction technologies (ARTs) with reference to the diagnosis and management of diseases affecting fertility. Our goal is to educate our readership in the translation of basic and clinical discoveries made from human or relevant animal models to the safe and efficacious practice of human ARTs. The scientific rigor and ethical standards embraced by the JARG editorial team ensures a broad international base of expertise guiding the marriage of contemporary clinical research paradigms with basic science discovery. JARG publishes original papers, minireviews, case reports, and opinion pieces often combined into special topic issues that will educate clinicians and scientists with interests in the mechanisms of human development that bear on the treatment of infertility and emerging innovations in human ARTs. The guiding principles of male and female reproductive health impacting pre- and post-conceptional viability and developmental potential are emphasized within the purview of human reproductive health in current and future generations of our species. The journal is published in cooperation with the American Society for Reproductive Medicine, an organization of more than 8,000 physicians, researchers, nurses, technicians and other professionals dedicated to advancing knowledge and expertise in reproductive biology.