Detection of the ABCA4 gene mutations using next-generation sequencing

Abstract

Purpose: To identify mutations in the ABCA4 gene in a group of patients with Stargardt disease, fundus flavimaculatus or cone-rod dystrophy

Methods: Genomic DNA isolated from peripheral blood of 58 patients served as a template. The introduced variant of next-generation sequencing is based on the preparation of an amplicon library containing all coding sequences of the ABCA4 gene (50 exons). Next, the amplicons were sequenced using the genomic sequencer GS Junior System (Roche).

Results: Analysis of the ABCA4 gene in a group of 58 patients enabled identification of 32 different known mutations and 20 different novel potentially pathogenic variants. Presence of these genetic changes has been confirmed by standard DNA sequencing. Our study enabled identification of the genetic cause of retinal disease in 88% of patients. Three patients (5%) did not carry any mutation in the ABCA4 gene and in four patients (7%) only one mutation was found.

Conclusions: Patients with an unknown cause of the retinal disease will be examined using whole genome sequencing.

Keywords: 539 genetics • 696 retinal degenerations: hereditary • 537 gene screening