IROme, a New High-Throughput Molecular Tool for the Diagnosis of Inherited Retinal Dystrophies

Abstract EN

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.