Next generation sequencing based molecular genetic analysis of a Chinese Han family with autosomal retinitis pigmentosa

Authors: Zhou Xiaomin,  Huang Hui,  Wang Ying,  Wu Jing,  Fan Ning,  Jiang Shanming,  Liu Xuyang
DOI: 10.3760/cma.j.issn.2095-0160.2015.08.006
Published 2015-08-10
Cite as Chin J Exp Ophthalmol, 2015,33(8): 699-703.

Abstract                              [Download PDF] [Read Full Text]

Background

Retinitis pigmeutosa (RP) is a progressive inheritance disease. It is characterized by highly genetical and phenotypical heterogeneity. With the rapid development of genomics, new methods are applied to the genetic screening of RP.

Objective

This study was to characterize the clinical features of a Chinese family with autosomal RP and to screen the candidate genes.

Methods

Twelve members from this family were included in the study. All participants underwent complete ophthalmologic examinations. Targeted-capture next generation sequencing (NGS) based molecular genetic analysis was performed on two patients of this RP family(Ⅱ5, Ⅱ7). The DNA sample from the two patients was separately sequenced using custom capture gene chip, which includes 59 retinal disease genes. The sequencing results were analyzed by bioinformatics technology. Identified variations were verified in the rest family members by PCR and Sanger sequencing. This study was approved by Ethic Committee of West China Hospital, and informed consent was obtained from the subjects.

Results

Four members of this family were diagnosed as RP, and the rest were asymptomatic. Missense mutation (c. 3065T>C, p. Phe1022Ser) in USH2A and missense mutation (c. 1699G>A, p. Ala1319Gly) in PDE6A were found in two patients (Ⅱ5 and Ⅱ7). The variants were not co-segregated with the phenotype of this family. The causative mutation was not found by the targeted-capture NGS based eye disease chip, but it ruled out a large number of candidate genes for RP.

Conclusions

Our study suggests that targeted-capture NGS based eye disease chip can quickly detect mutations in known RP genes. It can be a new applicable and efficient method for molecular genetic analysis of ocular disease.

Key words:

Retinitis pigmentosa; Gene; Next generation sequencing; Molecular genetic analysis; Pedigree

Contributor Information

Zhou Xiaomin
Ophthalmic Laboratories & Department of Medical Affairs, West China Hospital, Sichuan University, Chengdu 610041, China
Huang Hui
Wang Ying
Wu Jing
Fan Ning
Jiang Shanming
Liu Xuyang
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Updated: March 23, 2023 — 3:37 am