Capture-based high-coverage NGS: A powerful tool to uncover a wide spectrum of mutation types

Jing Wang, Hui Yu, Victor Wei Zhang, Xia Tian, Yanming Feng, Guoli Wang, Elizabeth Gorman, Hao Wang, Richard E. Lutz, Eric S. Schmitt, Sandra Peacock, Lee Jun Wong

Research output: Contribution to journalArticlepeer-review

26 Scopus citations


Purpose:Next-generation sequencing (NGS) has been widely applied to clinical diagnosis. Target-gene capture followed by deep sequencing provides unbiased enrichment of the target sequences, which not only accurately detects single-nucleotide variations (SNVs) and small insertion/deletions (indels) but also provides the opportunity for the identification of exonic copy-number variants (CNVs) and large genomic rearrangements.Method:Capture NGS has the ability to easily detect SNVs and small indels. However, genomic changes involving exonic deletions/duplications and chromosomal rearrangements require more careful analysis of captured NGS data. Misaligned raw sequence reads may be more than just bad data. Some mutations that are difficult to detect are filtered by the preset analytical parameters. "Loose" filtering and alignment conditions were used for thorough analysis of the misaligned NGS reads. Additionally, using an in-house algorithm, NGS coverage depth was thoroughly analyzed to detect CNVs.Results:Using real examples, this report underscores the importance of the accessibility to raw sequence data and manual review of suspicious sequence regions to avoid false-negative results in the clinical application of NGS. Assessment of the NGS raw data generated by the use of loose filtering parameters identified several sequence aberrations, including large indels and genomic rearrangements. Furthermore, NGS coverage depth analysis identified homozygous and heterozygous deletions involving single or multiple exons.Conclusion:Our results demonstrate the power of deep NGS in the simultaneous detection of point mutations and intragenic exonic deletion in one comprehensive step.

Original languageEnglish (US)
Pages (from-to)513-521
Number of pages9
JournalGenetics in Medicine
Issue number5
StatePublished - May 1 2016

ASJC Scopus subject areas

  • Genetics(clinical)


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