Journal article

Development of High-Throughput Clinical Testing of RPGR ORF15 Using a Large Inherited Retinal Dystrophy Cohort.

John PW Chiang, Tina M Lamey, Nicholas K Wang, Jie Duan, Wei Zhou, Terri L McLaren, Jennifer A Thompson, Jonathan Ruddle, John N De Roach

Investigative Ophthalmology & Visual Science | Published : 2018


Purpose: Mutations in the ORF15 region of RPGR account for approximately half of all X-linked retinitis pigmentosa cases. However, a robust high-throughput method for the detection of ORF15 mutations has yet to be validated. We set out to develop the first clinically validated next-generation sequencing (NGS) method for the detection of mutations in this difficult-to-sequence region, including test accuracy and coverage data. Methods: As part of a blind-test, 145 research samples, previously tested by Sanger sequencing, and 81 clinical samples were evaluated using NGS of long-range PCR products fragmented with Illumina's Nextera library preparation kit (method 1), or with Centrillion's OneTu..

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