title: IROme, a new high-throughput molecular tool for the diagnosis of inherited retinal dystrophies.
abstract: The molecular diagnosis of retinal dystrophies is difficult because of the very important number of genes implicated and is rarely helped by genotype-phenotype correlations. This prompted us to develop IROme, a custom designed in solution-based targeted exon capture assay (SeqCap EZ Choice library, Roche NimbleGen) for 60 retinitis pigmentosa-linked genes and three candidate genes (942 exons). Pyrosequencing was performed on a Roche 454 GS Junior benchtop high-throughput sequencing platform. In total, 23 patients affected by retinitis pigmentosa were analyzed. Per patient, 39.6 Mb were generated, and 1111 sequence variants were detected on average, at a median coverage of 17-fold. After data filtering and sequence variant prioritization, disease-causing mutations were identified in ABCA4, CNGB1, GUCY2D, PROM1, PRPF8, PRPF31, PRPH2, RHO, RP2, and TULP1 for twelve patients (55%), ten mutations having never been reported previously. Potential mutations were identified in 5 additional patients, and in only 6 patients no molecular diagnosis could be established (26%). In conclusion, targeted exon capture and next-generation sequencing are a valuable and efficient approach to identify disease-causing sequence variants in retinal dystrophies.
authors.affiliation: Institute for Research in Ophthalmology (IRO), Grand-Champsec 64, 1950 Sion, Switzerland. daniel.schorderet@irovision.ch
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Schorderet
authors.forename: Alexandra
authors.initials: A
authors.lastname: Iouranova
authors.forename: Tatiana
authors.initials: T
authors.lastname: Favez
authors.forename: Leila
authors.initials: L
authors.lastname: Tiab
authors.forename: Pascal
authors.initials: P
authors.lastname: Escher
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods; D005091:Exons; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012174:Retinitis Pigmentosa / Q000175:diagnosis* / Q000235:genetics*