多重连接依赖探针扩增
杜氏肌营养不良
外显子
无义突变
突变
基因复制
肌营养不良
遗传学
DNA测序
桑格测序
移码突变
基因
错义突变
生物
分子生物学
作者
Shuang Li,Ying Bai,Zhenhua Zhao,Xiangdong Kong
出处
期刊:PubMed
[National Institutes of Health]
日期:2016-12-10
卷期号:33 (6): 762-767
标识
DOI:10.3760/cma.j.issn.1003-9406.2016.06.004
摘要
OBJECTIVE: To perform mutation analysis for 81 unrelated patients with Duchenne/Becker muscular dystrophy (DMD/BMD) from Henan Province. METHODS: Multiplex ligation-dependent probe amplification (MLPA) was used to detect potential deletion/duplications of the DMD gene. Those with single exon deletions were validated with PCR amplification and Sanger sequencing to rule out false positive results. Patients with negative MLPA results were further analyzed with next-generation sequencing (NGS), and the result was validated by Sanger sequencing. RESULTS: DMD gene deletion/duplications were detected in 67 cases by MLPA, and exons 45-54 was the most frequently deleted. The phenotypes of 79.1% patients with a deletion or duplication has conformed to the reading frame rule. In addition, 13 mutations were detected by NGS and Sanger sequencing, which included 6 novel mutations including one frameshift mutation c.4708-4709insTG and 5 nonsense mutations (c.8812G>T, c.2131A>T, c.6035T>A, c.3426C>A, and c.3055C>T). CONCLUSION: This results have enriched the DMD gene mutation database. Combined MLPA, NGS and Sanger sequencing can greatly enhance the sensibility and specificity of genetic testing for the DMD/BMD.
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