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Abstract：
Retinitis pigmentosa (RP) is the most common inherited retinal dystrophy and a major cause of blindness. RP is caused by several variants of multiple genes, and genetic diagnosis by identifying these variants is important for optimizing treatment and estimating patient prognosis. Next-generation sequencing (NGS), which is currently widely used for diagnosis, is considered useful but is known to have limitations in detecting copy number variations (CNVs). In this study, we re-evaluated CNVs in EYS, the main causative gene of RP, identified via NGS using multiplex ligation-dependent probe amplification (MLPA). CNVs were identified in NGS samples of eight patients. To identify potential CNVs, MLPA was also performed on samples from 42 patients who were undiagnosed by NGS but carried one of the five major pathogenic variants reported in Japanese EYS-RP cases. All suspected CNVs based on NGS data in the eight patients were confirmed via MLPA. CNVs were found in 2 of the 42 NGS-undiagnosed RP cases. Furthermore, results showed that 121 of the 661 patients with RP had EYS as the causative gene, and 8.3% (10/121 patients with EYS-RP) had CNVs. Although NGS using the CNV calling criteria utilized in this study failed to identify CNVs in two cases, no false-positive results were detected. Collectively, these findings suggest that NGS is useful for CNV detection during clinical diagnosis of RP.
摘要：
视网膜色素变性（RP）是最常见的遗传性视网膜营养不良和失明的主要原因。RP是由多个基因的几种变异引起的，通过识别这些变异进行基因诊断对于优化治疗和估计患者预后很重要。下一代测序（NGS），目前广泛用于诊断，被认为是有用的,但已知在检测拷贝数变异(CNV)方面具有局限性。在这项研究中,我们在EYS中重新评估了CNVs，RP的主要致病基因，使用多重连接依赖性探针扩增(MLPA)通过NGS鉴定。在8名患者的NGS样品中鉴定了CNV。为了识别潜在的CNVs，还对来自42名患者的样本进行了MLPA，这些患者未被NGS诊断，但携带了日本EYS-RP病例中报道的五种主要致病变异之一。通过MLPA确认了8名患者中基于NGS数据的所有疑似CNV。在42例未诊断的NGSRP病例中，有2例发现了CNV。此外，结果显示，661例RP患者中有121例以EYS为致病基因，8.3%(10/121例EYS-RP患者)有CNVs。尽管NGS使用本研究中使用的CNV调用标准未能在两种情况下识别CNV，未检测到假阳性结果.总的来说，这些发现表明NGS在RP的临床诊断中对CNV的检测是有用的。
