Reproductive Medicine Center, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, Guangdong, China.
Guangdong Provincial Key Laboratory of Reproductive Medicine, Guangzhou, 510080, Guangdong, China.
J Assist Reprod Genet. 2020 Mar;37(3):549-557. doi: 10.1007/s10815-020-01732-7. Epub 2020 Mar 9.
To evaluate the efficacy of preimplantation genetic testing (PGT) for α- and β-double thalassemia combined with aneuploidy screening using next-generation sequencing (NGS).
An NGS-based PGT protocol was performed between 2017 and 2018 for twelve couples, each of which carried both α- and β-thalassemia mutations. Trophectoderm biopsy samples underwent whole-genome amplification using multiple displacement amplification (MDA), followed by NGS for thalassemia detection and aneuploidy screening. A selection of several informative single nucleotide polymorphisms (SNPs) established haplotypes. Aneuploidy screening was performed only on unaffected noncarriers and carriers. Unaffected and euploid embryos were transferred into the uterus through frozen-thawed embryo transfer (FET).
A total of 280 oocytes were retrieved following 18 ovum pick-up (OPU) cycles, with 182 normally fertilized and 112 cultured to become blastocysts. One hundred and seven (95.5%, 107/112) blastocysts received conclusive PGT results, showing 56 (52.3%, 56/107) were unaffected. Thirty-seven (66.1%, 37/56) of the unaffected were also identified as euploid. One family had no transferable embryos. Unaffected and euploid embryos were then transferred into the uterus of the other 11 couples resulting in 11 healthy live births. The clinical pregnancy rate was 61.1% (11/18) per OPU and 68.8% (11/16) per FET, with no miscarriage reported. Seven families accepted the prenatal diagnosis and received consistent results with the NGS-based PGT.
This study indicated that NGS could realize the simultaneous PGT of double thalassemia and aneuploidy screening in a reliable and accurate manner. Moreover, it eliminated the need for multiple biopsies, alleviating the potential damages to the pre-implanted blastocysts.
评估使用下一代测序（NGS）进行植入前遗传检测（PGT）α-和β-双重地中海贫血与非整倍体筛查的疗效。
2017 年至 2018 年期间，12 对夫妇进行了基于 NGS 的 PGT 方案，每对夫妇均携带α-和β-地中海贫血突变。滋养外胚层活检样本使用多重置换扩增（MDA）进行全基因组扩增，然后进行 NGS 检测地中海贫血和非整倍体筛查。选择了几个信息丰富的单核苷酸多态性（SNP）来建立单倍型。仅对未受影响的非携带者和携带者进行非整倍体筛查。通过冷冻胚胎移植（FET）将不受影响和整倍体胚胎转移到子宫中。
18 次卵母细胞采集（OPU）周期共获得 280 个卵母细胞，其中 182 个正常受精，112 个培养成囊胚。107 个（95.5%，107/112）囊胚获得了明确的 PGT 结果，其中 56 个（52.3%，56/107）不受影响。37 个（66.1%，37/56）不受影响的胚胎也被鉴定为整倍体。一个家庭没有可转移的胚胎。然后将不受影响和整倍体的胚胎转移到其他 11 对夫妇的子宫中，导致 11 例健康的活产。每 OPU 的临床妊娠率为 61.1%（11/18），每 FET 的妊娠率为 68.8%（11/16），无流产报告。7 个家庭接受了产前诊断，并获得了与 NGS 为基础的 PGT 一致的结果。
本研究表明，NGS 可以可靠、准确地同时进行双重地中海贫血和非整倍体筛查的 PGT。此外，它消除了对多个活检的需求，减轻了对植入前囊胚的潜在损害。
