title: Next-generation genetic testing for retinitis pigmentosa.
abstract: Molecular diagnostics for patients with retinitis pigmentosa (RP) has been hampered by extreme genetic and clinical heterogeneity, with 52 causative genes known to date. Here, we developed a comprehensive next-generation sequencing (NGS) approach for the clinical molecular diagnostics of RP. All known inherited retinal disease genes (n = 111) were captured and simultaneously analyzed using NGS in 100 RP patients without a molecular diagnosis. A systematic data analysis pipeline was developed and validated to prioritize and predict the pathogenicity of all genetic variants identified in each patient, which enabled us to reduce the number of potential pathogenic variants from approximately 1,200 to zero to nine per patient. Subsequent segregation analysis and in silico predictions of pathogenicity resulted in a molecular diagnosis in 36 RP patients, comprising 27 recessive, six dominant, and three X-linked cases. Intriguingly, De novo mutations were present in at least three out of 28 isolated cases with causative mutations. This study demonstrates the enormous potential and clinical utility of NGS in molecular diagnosis of genetically heterogeneous diseases such as RP. De novo dominant mutations appear to play a significant role in patients with isolated RP, having major implications for genetic counselling.
authors.affiliation: Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
authors.forename: Kornelia
authors.initials: K
authors.lastname: Neveling
authors.forename: Rob W J
authors.initials: RW
authors.lastname: Collin
authors.forename: Christian
authors.initials: C
authors.lastname: Gilissen
authors.forename: Ramon A C
authors.initials: RA
authors.lastname: van Huet
authors.forename: Linda
authors.initials: L
authors.lastname: Visser
authors.forename: Michael P
authors.initials: MP
authors.lastname: Kwint
authors.forename: Sabine J
authors.initials: SJ
authors.lastname: Gijsen
authors.forename: Marijke N
authors.initials: MN
authors.lastname: Zonneveld
authors.forename: Nienke
authors.initials: N
authors.lastname: Wieskamp
authors.forename: Joep
authors.initials: J
authors.lastname: de Ligt
authors.forename: Anna M
authors.initials: AM
authors.lastname: Siemiatkowska
authors.forename: Lies H
authors.initials: LH
authors.lastname: Hoefsloot
authors.forename: Michael F
authors.initials: MF
authors.lastname: Buckley
authors.forename: Ulrich
authors.initials: U
authors.lastname: Kellner
authors.forename: Kari E
authors.initials: KE
authors.lastname: Branham
authors.forename: Anneke I
authors.initials: AI
authors.lastname: den Hollander
authors.forename: Alexander
authors.initials: A
authors.lastname: Hoischen
authors.forename: Carel
authors.initials: C
authors.lastname: Hoyng
authors.forename: B Jeroen
authors.initials: BJ
authors.lastname: Klevering
authors.forename: L Ingeborgh
authors.initials: LI
authors.lastname: van den Born
authors.forename: Joris A
authors.initials: JA
authors.lastname: Veltman
authors.forename: Frans P M
authors.initials: FP
authors.lastname: Cremers
authors.forename: Hans
authors.initials: H
authors.lastname: Scheffer
mesh_terms: D000483:Alleles; D019295:Computational Biology / Q000379:methods; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008297:Male; D009154:Mutation; D010375:Pedigree; D010641:Phenotype; D011786:Quality Control; D015203:Reproducibility of Results; D012174:Retinitis Pigmentosa / Q000175:diagnosis* / Q000235:genetics