Genetics Working Group, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia.
Dr. Sardjito General Hospital, Yogyakarta, 55281, Indonesia.
F1000Res. 2023 Nov 17;11:148. doi: 10.12688/f1000research.73476.3. eCollection 2022.
Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations in the gene. The full mutation spectrum of the gene in Indonesian patients is currently unknown. Mutation-specific therapies are currently being developed, such as exon skipping or stop codon read-through therapy. This study was conducted with the aim of identifying the mutation spectrum of the gene in Indonesia to guide future development and application of feasible therapeutic strategies.
This study is a cross sectional study that enrolled 43 male patients with a clinical suspicion of or BMD. Multiplex ligation-dependent probe amplification (MLPA) reaction was performed to screen for the common mutations in the gene.
Out of 43 subjects, deletions accounted for 69.77% (n=30) cases, while duplications were found in 11.63% (n=5) cases. One novel duplication spanning exons 2 to 62 was identified. Deletion mutations clustered around the distal (66.67%) and proximal (26.67%) hot spot regions of the gene while duplication mutations were observed solely at the proximal region. Two false positive cases of single exon deletion detected through MLPA were attributed to sequence mutations affecting primer ligation sites, confirming the need to validate all single exon deletions when using this screening method. Analysis of available maternal DNA samples showed that the rate of de novo mutations (48.15%) appears higher than expected in this population. Out of 31 patients who were classified as DMD based on clinical and genotype characterizations, 60.47% (n=26) of cases were suitable for exon skipping therapy.
This is the first comprehensive study showing the feasibility of implementing the MLPA method for routine screening of patients in Indonesia. This is also the first study showing the potential applicability of exon skipping therapy in the majority of cases in the country.
杜氏肌营养不良症（DMD）和贝克肌营养不良症（BMD）是由 基因的突变引起的等位基因疾病。目前尚不清楚印度尼西亚患者 基因的完整突变谱。目前正在开发针对特定突变的疗法，例如外显子跳跃或终止密码子通读疗法。本研究旨在确定印度尼西亚 基因的突变谱，以指导未来可行治疗策略的开发和应用。
本研究是一项横断面研究，共纳入 43 名临床疑似 DMD 或 BMD 的男性患者。采用多重连接依赖性探针扩增（MLPA）反应筛选 基因中的常见突变。
在 43 名受试者中，缺失占 69.77%（n=30），而重复占 11.63%（n=5）。发现了一种跨越外显子 2 至 62 的新型重复。缺失突变聚集在 基因的远端（66.67%）和近端（26.67%）热点区域，而重复突变仅发生在近端区域。通过 MLPA 检测到的两个假阳性单外显子缺失病例归因于影响引物连接位点的序列突变，证实了在使用这种筛选方法时需要验证所有单外显子缺失。对可用的母体 DNA 样本进行分析表明，该人群中新生突变的发生率（48.15%）似乎高于预期。在根据临床和基因型特征分类为 DMD 的 31 名患者中，60.47%（n=26）的病例适合外显子跳跃治疗。
这是第一项全面研究，表明在印度尼西亚实施 MLPA 方法常规筛查 患者的可行性。这也是第一项表明外显子跳跃治疗在该国大多数 病例中具有潜在适用性的研究。
