利用目标基因捕获结合二代测序技术发现左心室心肌致密化不全MYBPC3基因错义突变  

Targeted next-generation sequencing in isolated left-ventricular noncompaction missense mutations in MYBPC3

作　　者：李然[1] 郭俊[1] 丁文虹[2] 焦萌[2] 李小燕[1] 王绿娅[1] 杜杰[1] 

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所,心血管重大疾病防治协同创新中心(2011协同创新中心)-省部共建心血管重塑相关疾病教育部重点实验室,100029 [2]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所,小儿心脏中心,100029

出　　处：《心肺血管病杂志》2016年第1期20-24,共5页Journal of Cardiovascular and Pulmonary Diseases

基　　金：北京心血管疾病协同创新研究中心基金(PXM2013_014226_07_000088)

摘　　要：目的:建立目标基因捕获结合第二代测序技术,对孤立性左心室心肌致密化不全(IVNC)患者的已知致病基因MYBPC3进行突变筛查。方法:收集5例IVNC患者及一级亲属的超声影像学资料,并提取外周血全基因组DNA,设计MYBPC3外显子区域特异性捕获探针,与基因组DNA文库进行杂交,富集目标基因组区域DNA片段,利用二代测序技术,确定突变位点,并使用Sanger测序法在其一代亲属中验证。结果:目标基因特异性捕获探针可有效地捕捉并富集基因组DNA目标靶片段。在5例IVNC患者中,发现1例MYBPC3基因杂合非同义突变c.G1000A(p.E334K),该突变位于MYBPC3基因第13外显子中,测序深度249.65。经过数据分析与Sanger测序验证后,父亲发现此突变位点,母亲未发现提示突变的父亲的遗传。结论:本研究所建立的Gen Cap目标基因捕获测序技术结合二代测序技术成功发现了MYBPC3基因突变。Objective： To establish a target gene capture combined with the second generation sequencing technology,we screened the Isolation Left Ventricular Noncompaction（ IVNC） patients for known pathogenic gene to investigate the relationship between the IVNC and MYBPC3 mutations. Methods： Collecting 5cases of ultrasonic imaging data of IVNC patients and their first-degree relatives by whole-genome DNA extracted from peripheral blood. Using Ilummina hiseq2000 second-generation sequencing,a corresponding exon region specific capture probe was designed. Genomic DNA library was hybridized and the target DNA genome area was enriched to identify the mutations. The Sanger sequencing validates the mutations in the first-degree of relatives. Results： The target gene specificity capture probe can effectively capture and enrich the genomic DNA targeted segments. In 5 patients with IVNC,we find one MYBPC3 gene heterozygous mutation,the gene mutations in the thirteen exons of MYBPC3,the average sequencing depth is 209. 5. After data analysis and the Sanger sequencing,a validate the new non-synonymous mutation c. G1000A（ p. E334K）. Conclusion： This research established Gen Cap capture target gene sequencing technology combined with the next-generation sequencing technology,we successfully found a new MYBPC3 gene mutations. This method is rapid and effective which enables us to have a better understanding on molecular etiology of IVNC.

关 键 词：孤立性左心室心肌致密化不全 MYBPC3基因 目标基因捕获技术 二代测序技术 

分 类 号：R54[医药卫生—心血管疾病]