Captured long-read sequecing provides an efficient and accurate method for molecular diagnosis of Duchenne and Becker muscular dystrophies

Abstract Background: Duchenne and Becker muscular dystrophies are not caused by deletions and duplications in the dystrophin (DMD) gene alone. A number of small and complex mutations in DMD were being found by multiple methods combined screening, including the next generation sequencing. However, there is still absent an effective method that could detect all types the potential variants in DMD. Therefore, in this study we explored a one-step detection method for DMD gene mutation based on long-read sequencing technology. Methods: A whole DMD gene panel including 20kb flanking sequences of the up and down stream of the DMD gene was designed. Pacific Biosciences and Oxford Nanopore Technologies were used to evaluate the capture and sequencing performance of the panel. A total of 129 subjects were selected for single-blind deep investigation and validation. Results: The results demonstrated that the long-read sequencing based DMD gene panel could integrally and accurately detect the multiple types of the variants in one-step. The noncontiguous variants were definitively corrected and attributed to translocation or inversion. Meanwhile, the micro insertion and deletion and the single nucleotide variants, especially the deep intronic variants, could be detected exactly compared with short-read sequencing technologies. Additionally, the captured long-read sequencing method could attain higher accuracy in female carrier mutation detection. Conclusion: This study illustrated that captured long-read sequencing could uncover the real features of DMD rearrangements via the effective junction reads analysis, and provide a complete and precise insight into the DMD gene mutation. Further, improve the molecular treatment of DMD/BMDin a base-pair resolution.