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This abstract is assigned to session BIO: Belgian Immuno Ophthalmology Club

Title

FRO: Development of a next-generation sequencing platform for retinal dystrophies, with LCA and RP as proof of concept.

Purpose	Leber Congenital Amaurosis (LCA) is a rare congenital retinal dystrophy associated with 16 genes. Recent breakthroughs in LCA gene therapy offer the first prospect of treating inherited blindness, which requires an unequivocal and early molecular diagnosis. While present genetic tests do not address this due to a tremendous genetic heterogeneity, massively parallel sequencing (MPS) strategies might bring a solution. Here, we developed a comprehensive molecular test for LCA based on targeted MPS of all exons of 16 known LCA genes.
Methods	We designed a unique and flexible workflow for targeted resequencing of all 236 exons from 16 LCA genes based on qPCR amplicon ligation, shearing and parallel sequencing of multiple patients on a single lane of a short read sequencer. Twenty-two pre-screened LCA patients were included, five of which with known molecular cause.
Results	Validation of 107 variations was performed as proof of concept. In addition, the causal genetic defect and a single heterozygous mutation were identified in 3 and 5 out of 17 patients without previously identified mutations, respectively.
Conclusion	We propose a novel targeted MPS-based approach that is suitable for accurate, fast and cost-effective early molecular testing in LCA, and easily applicable in other genetic disorders.

Author 1

Last name	COPPIETERS
Initials	F
Department	Center for Medical Genetics Ghent, Ghent University
City	Ghent