title: Capture-based high-coverage NGS: a powerful tool to uncover a wide spectrum of mutation types.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  Our results demonstrate the power of deep NGS in the simultaneous detection of point mutations and intragenic exonic deletion in one comprehensive step.Genet Med 18 5, 513-521.
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Jing
authors.identifier: 0000-0001-9178-7731
authors.initials: J
authors.lastname: Wang
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Yu
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Victor Wei
authors.initials: VW
authors.lastname: Zhang
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Tian
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Yanming
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Guoli
authors.initials: G
authors.lastname: Wang
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Gorman
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Munroe-Meyer Institute, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Richard E
authors.initials: RE
authors.lastname: Lutz
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Schmitt
authors.affiliation: Baylor Miraca Genetics Laboratories, Houston, Texas, USA.
authors.forename: Sandra
authors.identifier: 0000-0001-8119-3745
authors.initials: S
authors.lastname: Peacock
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Lee-Jun
authors.initials: LJ
authors.lastname: Wong
mesh_terms: D000465:Algorithms; D056915:DNA Copy Number Variations / Q000235:genetics; D005091:Exons; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D006579:Heterozygote; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods* / Q000639:trends; D006720:Homozygote; D006801:Humans; D054643:INDEL Mutation / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics