第二代测序技术在假肥大型肌营养不良基因诊断中的应用

摘要

[Objective]To detect genetic causes of Duchenne muscular dystrophy(DMD).[Methods] Next-generation sequencing was used to detect 6 DMD patients in whom no exonic deletions were detected by multiplex PCR.Sanger sequencing and multiplex ligation-dependent probe amplification were used to confirm the results.[Results] One case was found to have deletions of exons 10 and 11,1 had exons 16 and 17 duplication,4 cases have 8 point mutations including c.2776C＞T,c.5475delA,c.6391_6392delCA,IVS64+ 1G＞A,c.2645A＞G,c.5244G＞A,c.7728T＞C,c.8729A＞T,c.8734A＞G and c.8810G＞A.The former 4 mutations are auspicious pathogenicity,the other 6 mutations are polymorphism in population.Three novel mutations (IVS64+ 1G＞A,c.6391_6392delCA (p.Q2131NfsX3) and p.Q926X (CAG＞TAG) were not reported before.[Conclusion] Next-generation sequencing technology is a useful tool for the detection of deletion,duplication and point mutation,which is valuable for clinical application.%目的 对假肥大型肌营养不良患者进行基因检测.方法 采用目标区序列捕获及第2代高通量测序技术对6例假肥大型肌营养不良患者进行检测；采用第1代测序技术、多重连接依赖的探针扩增技术(multiplex ligation-dependent probe amplification,MLPA)对患者及其母亲的基因型进行验证.结果 1例为第10和11外显子缺失,1例为第16和17外显子重复,4例为点突变.共发现10种变异:c.2776C＞T、c.5475delA、c.6391_ 6392delCA、IVS64+1G＞A、c.2645A＞G、c.5244G＞A、c.7728T＞C、c.8729A＞T、e.8734A＞G和c.8810G＞A,前4种为可疑致病性变异,后6种为人群中的多态.其中3例为首次发现的新突变(IVS64+1G＞A、c.6391_6392delCA(p.Q2131NfsX3)和p.Q926X(CAG＞TAG).结论 通过第2代测序技术可以在一个反应中准确检测出DMD基因的缺失、重复和点突变,具有一定的临床应用价值.