Clinical application of nanopore sequencing for haplotype linkage analysis in preimplantation genetic testing for Duchenne muscular dystrophy

Abstract

Abstract Background Duchenne muscular dystrophy (DMD) is an X-linked recessive disease that affects approximately 1 in 5,000 newborn males. Most patients die of cardiac failure and/or respiratory insufficiency at approximately 20–40 years of age. Preimplantation genetic testing for monogenic disorders (PGT-M) has been successfully used to help couples with DMD mutations produce an unaffected offspring. However, the current PGT-M based on next-generation sequencing (NGS) cannot provide an effective PGT technique for DMD that involves de novo mutations or an incomplete pedigree. This study assessed the feasibility of using nanopore sequencing in PGT-M for DMD. The accuracy of the nanopore sequencing results was confirmed using an NGS-based method, which was further validated through amniocentesis. Results Using nanopore sequencing, we successfully determined the breakpoints of the DMD mutation and identified a set of informative heterozygous single-nucleotide polymorphisms (SNPs) in the maternal carrier. Subsequently, we conducted a haplotype linkage analysis with embryonic SNPs without the need for additional family members and successfully identified a disease-free euploid embryo. These results were consistent with the findings obtained using NGS and aligned with the results of amniocentesis. Conclusions Our results suggest that nanopore sequencing is a potential tool for preimplantation haplotype linkage analysis in PGT-M for DMD, particularly in families lacking probands. This finding is essential for nanopore sequencing to aid in reducing the propagation of DMD in the population.