title: Next-generation-based targeted sequencing as an efficient tool for the study of the genetic background in Hirschsprung patients.
abstract: BACKGROUND
  The development of next-generation sequencing (NGS) technologies has a great impact in the human variation detection given their high-throughput. These techniques are particularly helpful for the evaluation of the genetic background in disorders of complex genetic etiology such as Hirschsprung disease (HSCR). The purpose of this study was the design of a panel of HSCR associated genes as a rapid and efficient tool to perform genetic screening in a series of patients.
  
  
  METHODS
  We have performed NGS-based targeted sequencing (454-GS Junior) using a panel containing 26 associated or candidate genes for HSCR in a group of 11 selected HSCR patients.
  
  
  RESULTS
  The average percentage of covered bases was of 97%, the 91.4% of the targeted bases were covered with depth above 20X and the mean coverage was 422X. In addition, we have found a total of 13 new coding variants and 11 new variants within regulatory regions among our patients. These outcomes allowed us to re-evaluate the genetic component associated to HSCR in these patients.
  
  
  CONCLUSIONS
  Our validated NGS panel constitutes an optimum method for the identification of new variants in our patients. This approach could be used for a fast, reliable and more thorough genetic screening in future series of patients.
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Seville, Spain. berta.luzon.exts@juntadeandalucia.es.|Centre for Biomedical Network Research on Rare Diseases (CIBERER), Seville, Spain. berta.luzon.exts@juntadeandalucia.es.
authors.forename: Berta
authors.initials: B
authors.lastname: Luzón-Toro
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Seville, Spain. laura.espino.exts@juntadeandalucia.es.|Centre for Biomedical Network Research on Rare Diseases (CIBERER), Seville, Spain. laura.espino.exts@juntadeandalucia.es.
authors.forename: Laura
authors.initials: L
authors.lastname: Espino-Paisán
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Seville, Spain. raquelm.fernandez.sspa@juntadeandalucia.es.|Centre for Biomedical Network Research on Rare Diseases (CIBERER), Seville, Spain. raquelm.fernandez.sspa@juntadeandalucia.es.
authors.forename: Raquel Ma
authors.initials: RM
authors.lastname: Fernández
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Seville, Spain. m.martin.s@hotmail.com.|Centre for Biomedical Network Research on Rare Diseases (CIBERER), Seville, Spain. m.martin.s@hotmail.com.
authors.forename: Marta
authors.initials: M
authors.lastname: Martín-Sánchez
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Seville, Spain. guillermo.antinolo.sspa@juntadeandalucia.es.|Centre for Biomedical Network Research on Rare Diseases (CIBERER), Seville, Spain. guillermo.antinolo.sspa@juntadeandalucia.es.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Antiñolo
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Seville, Spain. salud.borrego.sspa@juntadeandalucia.es.|Centre for Biomedical Network Research on Rare Diseases (CIBERER), Seville, Spain. salud.borrego.sspa@juntadeandalucia.es.
authors.forename: Salud
authors.initials: S
authors.lastname: Borrego
mesh_terms: D001483:Base Sequence; D019295:Computational Biology; D005260:Female; D015723:Gene Library; D005820:Genetic Testing / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006627:Hirschsprung Disease / Q000235:genetics* / Q000473:pathology; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010641:Phenotype; D012680:Sensitivity and Specificity; D013030:Spain