title: Panel-based next generation sequencing as a reliable and efficient technique to detect mutations in unselected patients with retinal dystrophies.
abstract: Hereditary retinal dystrophies (RD) constitute a group of blinding diseases that are characterized by clinical variability and pronounced genetic heterogeneity. The different forms of RD can be caused by mutations in >100 genes, including >1600 exons. Consequently, next generation sequencing (NGS) technologies are among the most promising approaches to identify mutations in RD. So far, NGS is not routinely used in gene diagnostics. We developed a diagnostic NGS pipeline to identify mutations in 170 genetically and clinically unselected RD patients. NGS was applied to 105 RD-associated genes. Underrepresented regions were examined by Sanger sequencing. The NGS approach was successfully established using cases with known sequence alterations. Depending on the initial clinical diagnosis, we identified likely causative mutations in 55% of retinitis pigmentosa and 80% of Bardet-Biedl or Usher syndrome cases. Seventy-one novel mutations in 40 genes were newly associated with RD. The genes USH2A, EYS, ABCA4, and RHO were more frequently affected than others. Occasionally, cases carried mutations in more than one RD-associated gene. In addition, we found possible dominant de-novo mutations in cases with sporadic RD, which implies consequences for counseling of patients and families. NGS-based mutation analyses are reliable and cost-efficient approaches in gene diagnostics of genetically heterogeneous diseases like RD.
authors.affiliation: CeGaT GmbH, Tübingen, Germany.
authors.forename: Nicola
authors.initials: N
authors.lastname: Glöckle
authors.affiliation: Molecular Genetics Laboratory, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Kohl
authors.affiliation: CeGaT GmbH, Tübingen, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Mohr
authors.affiliation: CeGaT GmbH, Tübingen, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Scheurenbrand
authors.affiliation: CeGaT GmbH, Tübingen, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Sprecher
authors.affiliation: Molecular Genetics Laboratory, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Weisschuh
authors.affiliation: Centre for Ophthalmology, University of Tübingen, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Bernd
authors.affiliation: University Eye Hospital, Ludwigs-Maximilians-University, Munich, Germany.
authors.forename: Günther
authors.initials: G
authors.lastname: Rudolph
authors.affiliation: CeGaT GmbH, Tübingen, Germany.
authors.forename: Max
authors.initials: M
authors.lastname: Schubach
authors.affiliation: Department of Ophthalmology, University of Freiburg, Freiburg, Germany.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Poloschek
authors.affiliation: Centre for Ophthalmology, University of Tübingen, Germany.
authors.forename: Eberhart
authors.initials: E
authors.lastname: Zrenner
authors.affiliation: CeGaT GmbH, Tübingen, Germany.
authors.forename: Saskia
authors.initials: S
authors.lastname: Biskup
authors.affiliation: 1] University of Zurich, Institute of Medical Molecular Genetics, Schwerzenbach, Switzerland [2] Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland [3] Neuroscience Center Zurich (ZNZ), University and ETH Zurich, Zurich, Switzerland.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Berger
authors.affiliation: Molecular Genetics Laboratory, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Wissinger
authors.affiliation: University of Zurich, Institute of Medical Molecular Genetics, Schwerzenbach, Switzerland.
authors.forename: John
authors.initials: J
authors.lastname: Neidhardt
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D005091:Exons; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics; D020022:Genetic Predisposition to Disease; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D009154:Mutation; D057089:Pathology, Molecular; D010375:Pedigree; D058499:Retinal Dystrophies / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D012174:Retinitis Pigmentosa / Q000209:etiology / Q000235:genetics*; D017422:Sequence Analysis, DNA; D052245:Usher Syndromes / Q000209:etiology / Q000235:genetics* / Q000473:pathology
chemical_list: C056475:ABCA4 protein, human; D018528:ATP-Binding Cassette Transporters; C533105:EYS protein, human; D016326:Extracellular Matrix Proteins; D005136:Eye Proteins; C112735:USH2A protein, human