Improved Diagnosis of Inherited Retinal Dystrophies by High-Fidelity PCR of ORF15 followed by Next-Generation Sequencing

Retinitis pigmentosa (RP) is the most common form of retinal dystrophy. The disease is characterized by the progressive degeneration of photoreceptors, ultimately leading to blindness. The exon ORF15 of RP  GTPase regulator (RPGR) is a mutation hot spot for X-linked RP and one form of cone dystrophy. However, accurate molecular testing of ORF15 is challenging because of a large segment of highly repetitive purine-rich sequence in this exon. ORF15 performs poorly in next-generation sequencing–base d panels or whole exome sequencing analysis, whereas Sanger sequencing of ORF15 requires special reagents and PCR conditions with multiple pairs of overlapping primers that often do not provide a clean sequence.

Source: Journal of Molecular Diagnostics - September 11, 2016 Category: Pathology Authors: Jianli Li, Jia Tang, Yanming Feng, Mingchu Xu, Rui Chen, Xuan Zou, Ruifang Sui, Emmanuel Y. Chang, Richard A. Lewis, Victor W. Zhang, Jing Wang, Lee-Jun C. Wong Tags: Technical Advance Source Type: research