A next-generation sequencing gene panel (MiamiOtoGenes) for comprehensive analysis of deafness genes.
A next-generation sequencing gene panel (MiamiOtoGenes) for comprehensive analysis of deafness genes.
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DOI：
10.1016/j.heares.2016.01.018
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发表时间：
2016-03
期刊：
影响因子：
2.8
通讯作者：
Liu X
中科院分区：
文献类型：
--
作者：
Tekin D;Yan D;Bademci G;Feng Y;Guo S;Foster J 2nd;Blanton S;Tekin M;Liu X
极端的遗传异质性沿着不同种族间致病变异体分布的显著差异使得单基因检测对听力损失的检测效率低下。我们开发了一个定制的捕获/下一代测序基因面板，包含146个已知的耳聋基因，总靶大小约为1 MB。这些基因是通过搜索数据库确定的，包括遗传性听力损失主页，人类基因组突变数据库（HGMD），在线孟德尔遗传人类（OMIM）和最近的同行评议的出版物有关的遗传学耳聋。设计涵盖所有编码外显子、UTR和每个外显子的25个碱基的内含子侧翼序列。为了验证我们的小组，我们使用了6个已知耳聋基因变异的阳性对照和8个来自听力损失个体的未解决样本。靶向外显子的平均覆盖率为697 X。平均而言，每个样品分别具有99.8%、96.2%和92.7%的1X、50 X和100 X读段的靶区域覆盖率。分析检测到了所有已知的核基因变异。这些结果证明了自定义捕获实验的准确性和可靠性。
Extreme genetic heterogeneity along with remarkable variation in the distribution of causative variants across in different ethnicities makes single gene testing inefficient for hearing loss. We developed a custom capture/next-generation sequencing gene panel of 146 known deafness genes with a total target size of approximately 1MB. The genes were identified by searching databases including Hereditary Hearing Loss Homepage, the Human Genome Mutation Database (HGMD), Online Mendelian Inheritance in Man (OMIM) and most recent peer-reviewed publications related to the genetics of deafness. The design covered all coding exons, UTRs and 25 bases of intronic flanking sequences for each exon. To validate our panel, we used 6 positive controls with variants in known deafness genes and 8 unsolved samples from individuals with hearing loss. Mean coverage of the targeted exons was 697X. On average, each sample had 99.8%, 96.2% and 92.7% of the targeted region coverage of 1X, 50X and 100X reads, respectively. Analysis detected all known variants in nuclear genes. These results prove the accuracy and reliability of the custom capture experiment.