New Lung Cancer Panel for High-Throughput Targeted Resequencing

Genomics & Informatics

Kyusang Lee¹  Jongsun Park²  Byung Chul Kim²  Sunghoon Lee³  Eun-Hye Kim³  Jong Bhak³ 

[1] Clinomics Inc., Seoul 138-961, Korea.;Personal Genomics Institute, Genome Research Foundation, AICT, Suwon 443-270, Korea.;Theragen Bio Institute, AICT, Suwon 443-270, Korea.;

关键词: high-throughput nucleotide sequencing;    lung neoplasms;    next-generation sequencing;    selector technology;    somatic mutation screening;    target enrichment;   

DOI  :  10.5808/GI.2014.12.2.50

来源: DOAJ

【 摘 要 】

We present a new next-generation sequencing-based method to identify somatic mutations of lung cancer. It is a comprehensive mutation profiling protocol to detect somatic mutations in 30 genes found frequently in lung adenocarcinoma. The total length of the target regions is 107 kb, and a capture assay was designed to cover 99% of it. This method exhibited about 97% mean coverage at 30× sequencing depth and 42% average specificity when sequencing of more than 3.25 Gb was carried out for the normal sample. We discovered 513 variations from targeted exome sequencing of lung cancer cells, which is 3.9-fold higher than in the normal sample. The variations in cancer cells included previously reported somatic mutations in the COSMIC database, such as variations in TP53, KRAS, and STK11 of sample H-23 and in EGFR of sample H-1650, especially with more than 1,000× coverage. Among the somatic mutations, up to 91% of single nucleotide polymorphisms from the two cancer samples were validated by DNA microarray-based genotyping. Our results demonstrated the feasibility of high-throughput mutation profiling with lung adenocarcinoma samples, and the profiling method can be used as a robust and effective protocol for somatic variant screening.

【 授权许可】

Unknown