Targeted DNA-HiSeq
Identification of Sequence Variants in Genetic Disease Genes using Targeted Next-Generation Sequencing
Identification of gene variants plays important roles on the research and diagnosis of genetic diseases. A combination of enrichment of targeted genes and next-generation sequencing (Targeted DNA-HiSeq) has high efficiency and low cost for targeted sequencing of interesting genes. To identify mutations associated with kinds of genetic diseases, we designed an array-based gene chip to capture all exons of 193 genes involving in 103 genetic diseases. To evaluate the accuracy, we selected seven samples from seven patients with six different genetic diseases involving into six disease-causing genes, and 103 samples from adult normal persons as control. Our resulting data showed that on average 99.154% of 3382 exons with 30-fold coverage were successfully detected using the Targeted DNA-HiSeq technology, and we found six known variants in four disease-causing genes and two novel mutations in other two disease-causing genes (STS gene for XLI, FBN1 gene for MFS). The results were then confimed by using  the Sanger sequencing. In conclusion, the targeted DNA-HiSeq combines next-generation sequencing with capture of sequences from a relevant subset of "high interest" genes. Testing by capturing sequences from a DNA library by hybridization to oligonucleotide probes specific for 193 genetic-disorder-related genes, this method showed high selectivity, improved mutation detection enabling discovery of novel variants, and provided more indel data. Thus, targeted DNA-HiSeq can be used to analyze the gene variant profiling of some monogenic diseases with some features of high sensitivity, high fidelity, high throughput and high speed.