Rapid and cost-effective molecular diagnosis using exome sequencing of one proband with autosomal dominant congenital cataract
Rapid and cost-effective molecular diagnosis using exome sequencing of one proband with autosomal dominant congenital cataract
复制标题
使用外显子组测序对一名患有常染色体显性先天性白内障的先证者进行快速且经济有效的分子诊断。
DOI：
10.1038/eye.2014.158
复制
发表时间：
2014-12-01
期刊：
影响因子：
3.9
通讯作者：
Zhang, M.
中科院分区：
文献类型：
--
作者：
Chen, J-H;Qiu, J.;Zhang, M.
由于遗传异质性高，利用常规候选基因筛查排除常染色体显性先天性白内障（ADCC）的已知突变和绘制新突变需要繁重的实验室工作。我们试图使用一种经济有效的外显子组测序策略来识别ADCC家系中的致病突变。方法选取1例核性白内障ADCC家系和200例无亲缘关系的老年性白内障对照，进行眼科综合检查。使用Illumina TruSeq exome Enrichment Kit捕获该家族先证者的全外显子组，然后使用Illumina HiSeq 2000测序仪进行测序。通过直接测序进行验证。结果整个外显子组，包括已知ADCC致病基因的所有外显子，筛选可能的致病突变。在核性白内障先证者的间隙连接蛋白α 8基因（GJA8）外显子2中发现了一个复发性错义突变c.773C >t （p. S258F）。结果经直接测序证实。突变在家族中显示出与疾病表型完全共分离，但在不相关的未受影响的对照中未观察到。结论通过成功测序一个先显子的全外显子组，并在一个ADCC家系中发现GJA8突变，本研究表明外显子组测序可作为一种快速、可靠、经济的方法用于ADCC的临床诊断和致病基因发现。
Purpose Due to high genetic heterogeneity, to exclude known mutations and map novel mutations in autosomal dominant congenital cataract (ADCC) using conventional candidate gene screening requires laborious laboratory work. We attempted to use a cost-effective exome sequencing strategy to identify disease-causing mutations in an ADCC pedigree.Methods An ADCC pedigree affected by nuclear cataract and 200 unrelated senile cataract controls were recruited and given comprehensive ophthalmic examination. Whole exome of the proband of the family was captured by the Illumina TruSeq Exome Enrichment Kit, followed by sequencing using Illumina HiSeq 2000 sequencer. Validation was performed by direct sequencing.Results The whole exome, including all exons of known ADCC disease-causing genes, was screened for possible disease-causing mutations. A recurrent missense mutation c.773C>T (p. S258F) in exon 2 of the gap junction protein alpha 8 gene (GJA8) was identified in the proband with nuclear cataract. The result was confirmed by direct sequencing. The mutation showed complete co-segregation with the disease phenotype in the family but was not observed in unrelated unaffected controls.Conclusion By successfully sequencing whole exome of only one proband and identifying a GJA8 mutation in one ADCC pedigree, the current study demonstrated that exome sequencing could serve as a rapid, robust, and cost-effective approach in clinical diagnosis and disease-causing gene discovery for ADCC.