Hiroki Kurahashi

Objective Xq chromosome duplication with complex rearrangements is generally acknowledged to be associated with neurodevelopmental disorders, such as Pelizaeus‐Merzbacher disease (PMD) and MECP2 duplication syndrome. For couples who required a PGT‐M (pre‐implantation genetic testing for monogenic disease) for these disorders, junction‐specific PCR is useful to directly detect pathogenic variants. Therefore, pre‐clinical workup for PGT‐M requires the identification of the junction of duplicated segments in PMD and MECP2 duplication syndrome, which is generally difficult. Methods In this report, we used nanopore long‐read sequencing targeting the X chromosome using an adaptive sampling method to identify breakpoint junctions in disease‐causing triplications. Results By long‐read sequencing, we successfully identified breakpoint junctions in one PMD case with PLP1 triplication and in another MECP2 triplication case in a single sequencing run. Surprisingly, the duplicated region involving MECP2 was inserted 45 Mb proximal to the original position. This inserted region was confirmed by FISH analysis. With the help of precise mapping of the pathogenic variant, we successfully re‐established STR haplotyping for PGT‐M and avoided any potential misinterpretation of the pathogenic allele due to recombination. Conclusion Long‐read sequencing with adaptive sampling in a PGT‐M pre‐clinical workup is a beneficial method for identifying junctions of chromosomal complex structural rearrangements.