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. 2014 Jan;22(1):99-104.
doi: 10.1038/ejhg.2013.72. Epub 2013 Apr 17.

Panel-based next generation sequencing as a reliable and efficient technique to detect mutations in unselected patients with retinal dystrophies

Nicola Glöckle  1 , Susanne Kohl  2 , Julia Mohr  1 , Tim Scheurenbrand  1 , Andrea Sprecher  1 , Nicole Weisschuh  2 , Antje Bernd  3 , Günther Rudolph  4 , Max Schubach  1 , Charlotte Poloschek  5 , Eberhart Zrenner  3 , Saskia Biskup  1 , Wolfgang Berger  6 , Bernd Wissinger  2 , John Neidhardt  7
Affiliations

Panel-based next generation sequencing as a reliable and efficient technique to detect mutations in unselected patients with retinal dystrophies

Nicola Glöckle et al. Eur J Hum Genet. 2014 Jan.

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.

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Figures

Figure 1
Figure 1
The NGS strategy applied to identify mutations in patients suffering from hereditary retinal dystrophy. The flow chart illustrates the main steps in the working procedure from analysis of the patient sample to assembly of a medical report.
Figure 2
Figure 2
Mutations identified in 170 patients affected by hereditary retinal dystrophies. (a) Different types of mutations and their frequencies. (b) Affected genes and their frequencies. The frequencies are shown in brackets. The lists summarize genes that were found to be affected once or twice.
See this image and copyright information in PMC

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