Xia Qiuping, Chang Tianli, Ding Taoli, Liu Zhen, Liu Jiaqi, Li Yanping, Yao Zhongyuan
Reproductive Medicine Center, Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China.
Clinical Research Center for Women's Reproductive Health in Hunan Province, Changsha, 410008, Hunan, China.
Sci Rep. 2025 Aug 20;15(1):30498. doi: 10.1038/s41598-025-16358-x.
Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive muscular dystrophy, ultimately leading to paralysis and death from cardiac failure and/or respiratory insufficiency. Preimplantation genetic testing for monogenic disorders (PGT-M) has proven effective in helping families with complete pedigrees carrying DMD mutations to produce unaffected offspring. This study was designed to evaluate the feasibility of using nanopore sequencing as an effective PGT-M technique for DMD, particularly in cases involving de novo mutations or incomplete pedigrees that cannot be analyzed using second-generation sequencing (SGS) alone. Nanopore sequencing was performed on two DMD female carriers. The precise breakpoints of the DMD mutations were detected with nanopore sequencing, and haplotypes were constructed based on flanking single-nucleotide polymorphisms (SNPs). Haplotype linkage analyses were subsequently performed by comparing parental SNPs with embryonic SNPs to determine whether the embryos inherited the maternal DMD mutation-carrying chromosome. We successfully identified disease-free euploid embryos for both pedigrees. These results were consistent with the data obtained using SGS and amniocentesis. Our results establish nanopore sequencing as a clinically applicable methodology for preimplantation haplotype linkage analysis in PGT-M for DMD, particularly valuable for families with non-informative pedigrees where traditional linkage analyses are not feasible. This finding is crucial for reducing the propagation of DMD in the population through the application of nanopore sequencing.
杜氏肌营养不良症（DMD）是一种X连锁隐性疾病，其特征为进行性肌营养不良，最终导致因心力衰竭和/或呼吸功能不全而瘫痪和死亡。单基因疾病植入前基因检测（PGT-M）已被证明可有效帮助携带DMD突变的完整家系的家庭生育未受影响的后代。本研究旨在评估使用纳米孔测序作为DMD的有效PGT-M技术的可行性，特别是在涉及新发突变或无法仅使用二代测序（SGS）分析的不完整家系的情况下。对两名DMD女性携带者进行了纳米孔测序。通过纳米孔测序检测到DMD突变的精确断点，并基于侧翼单核苷酸多态性（SNP）构建单倍型。随后通过将亲代SNP与胚胎SNP进行比较来进行单倍型连锁分析，以确定胚胎是否继承了携带母体DMD突变的染色体。我们成功为两个家系鉴定出无病整倍体胚胎。这些结果与使用SGS和羊膜穿刺术获得的数据一致。我们的结果确立了纳米孔测序作为PGT-M中DMD植入前单倍型连锁分析的临床适用方法，对于传统连锁分析不可行的无信息家系的家庭尤为有价值。这一发现对于通过应用纳米孔测序减少DMD在人群中的传播至关重要。
