Department of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, Spain.
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), 41013 Seville, Spain.
Int J Mol Sci. 2020 Dec 8;21(24):9355. doi: 10.3390/ijms21249355.
The management of unsolved inherited retinal dystrophies (IRD) cases is challenging since no standard pipelines have been established. This study aimed to define a diagnostic algorithm useful for the diagnostic routine and to address unsolved cases. Here, we applied a Next-Generation Sequencing-based workflow, including a first step of panel sequencing (PS) followed by clinical-exome sequencing (CES) and whole-exome sequencing (WES), in 46 IRD patients belonging to 42 families. Twenty-six likely causal variants in retinal genes were found by PS and CES. CES and WES allowed proposing two novel candidate ( and a X-linked region including ), both abundantly expressed in human retina according to RT-PCR and immunohistochemistry. After comparison studies, PS showed the best quality and cost values, CES and WES involved similar analytical efforts and WES presented the highest diagnostic yield. These results reinforce the relevance of panels as a first step in the diagnostic routine and suggest WES as the next strategy for unsolved cases, reserving CES for the simultaneous study of multiple conditions. Standardizing this algorithm would enhance the efficiency and equity of clinical genetics practice. Furthermore, the identified candidate genes could contribute to increase the diagnostic yield and expand the mutational spectrum in these disorders.
未解决的遗传性视网膜疾病（IRD）病例的管理具有挑战性，因为尚未建立标准的处理流程。本研究旨在定义一种可用于诊断常规并解决未解决病例的诊断算法。在这里，我们对 46 名属于 42 个家族的 IRD 患者应用了基于下一代测序的工作流程，包括首先进行面板测序（PS），然后进行临床外显子组测序（CES）和全外显子组测序（WES）。通过 PS 和 CES 发现了 26 个视网膜基因中的可能致病变异。CES 和 WES 提出了两个新的候选基因（和一个包含 X 连锁区域的），根据 RT-PCR 和免疫组织化学，这两个基因在人视网膜中均大量表达。经过比较研究，PS 显示出最佳的质量和成本价值，CES 和 WES 涉及相似的分析工作量，而 WES 则具有最高的诊断率。这些结果证实了将面板作为诊断常规的第一步的重要性，并建议将 WES 作为未解决病例的下一步策略，将 CES 保留用于同时研究多种情况。标准化该算法将提高临床遗传学实践的效率和公平性。此外，所鉴定的候选基因可能有助于提高这些疾病的诊断率并扩大突变谱。
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