BGI-Shenzhen, Shenzhen, China.
School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, China.
PLoS One. 2018 Apr 11;13(4):e0185237. doi: 10.1371/journal.pone.0185237. eCollection 2018.
Inherited eye diseases are major causes of vision loss in both children and adults. Inherited eye diseases are characterized by clinical variability and pronounced genetic heterogeneity. Genetic testing may provide an accurate diagnosis for ophthalmic genetic disorders and allow gene therapy for specific diseases.
A targeted gene capture panel was designed to capture exons of 283 inherited eye disease genes including 58 known causative retinitis pigmentosa (RP) genes. 180 samples were tested with this panel, 68 were previously tested by Sanger sequencing. Systematic evaluation of our method and comprehensive molecular diagnosis were carried on 99 RP patients.
96.85% targeted regions were covered by at least 20 folds, the accuracy of variants detection was 99.994%. In 4 of the 68 samples previously tested by Sanger sequencing, mutations of other diseases not consisting with the clinical diagnosis were detected by next-generation sequencing (NGS) not Sanger. Among the 99 RP patients, 64 (64.6%) were detected with pathogenic mutations, while in 3 patients, it was inconsistent between molecular diagnosis and their initial clinical diagnosis. After revisiting, one patient's clinical diagnosis was reclassified. In addition, 3 patients were found carrying large deletions.
We have systematically evaluated our method and compared it with Sanger sequencing, and have identified a large number of novel mutations in a cohort of 99 RP patients. The results showed a sufficient accuracy of our method and suggested the importance of molecular diagnosis in clinical diagnosis.
遗传性眼病是儿童和成人视力丧失的主要原因。遗传性眼病的特点是临床表现的变异性和明显的遗传异质性。基因检测可为眼科遗传疾病提供准确的诊断，并允许针对特定疾病进行基因治疗。
设计了一个靶向基因捕获面板，用于捕获包括 58 个已知致盲性色素性视网膜炎（RP）基因在内的 283 个遗传性眼病基因的外显子。该面板对 180 个样本进行了测试，其中 68 个样本之前已通过 Sanger 测序进行了测试。对 99 例 RP 患者进行了系统的方法评估和全面的分子诊断。
至少 20 倍覆盖了 96.85%的靶向区域，变异检测的准确率为 99.994%。在之前通过 Sanger 测序测试的 68 个样本中，有 4 个通过下一代测序（NGS）而非 Sanger 检测到了与临床诊断不一致的其他疾病的突变。在 99 例 RP 患者中，64 例（64.6%）检测到致病性突变，而在 3 例患者中，分子诊断与初始临床诊断不一致。重新检查后，1 例患者的临床诊断被重新分类。此外，还发现 3 例患者携带大片段缺失。
我们系统地评估了我们的方法，并将其与 Sanger 测序进行了比较，在 99 例 RP 患者的队列中鉴定了大量新的突变。结果表明我们的方法具有足够的准确性，并提示分子诊断在临床诊断中的重要性。
