METHODS: Patients meeting the diagnosis criteria of PKD with negative results of PRRT2-Sanger sequencing and WES were included in this study. Mutation screening and targeted high-throughput sequencing were performed to analyze and verify the sequencing results of the potential mutations.
RESULTS: Six patients with PKD with high mutation ratios of c.649dupC were screened using our targeted high-throughput sequencing from 26 PKD patients with negative results of PRRT2-Sanger sequencing and WES (frequency = 23.1%), which compensated for the comparatively shallow sequencing depth and statistical flaws in this region. Compared with the local normal population and other patients with PKD, the mutation ratios of c.649dupC of these six patients with PKD were much higher and also had truncated protein structures and differentially altered mRNA expression.
CONCLUSIONS: Based on the above studies, we emphasize the routine targeted high-throughput sequencing of the c.649 site in the PRRT2 gene in so-called genetic-testing-negative patients with PKD, and manually calculate the deletion and duplication mutations depth and ratios to lower the rate of clinical misdiagnosis.
方法:符合PKD诊断标准且PRRT2-Sanger测序和WES结果阴性的患者纳入本研究。进行突变筛选和靶向高通量测序以分析和验证潜在突变的测序结果。
结果:使用我们的靶向高通量测序从26例PRRT2-Sanger测序和WES结果阴性的PKD患者中筛选出6例具有c.649dupC高突变率的PKD患者(频率=23.1%),这弥补了该区域相对较浅的测序深度和统计缺陷。与当地正常人群和其他PKD患者相比,这6例PKD患者的c.649dupC突变率要高得多,蛋白结构截短,mRNA表达差异改变.
结论:基于上述研究,我们强调在所谓的遗传检测阴性PKD患者中对PRRT2基因中c.649位点进行常规的靶向高通量测序,并手动计算缺失和重复突变的深度和比率,以降低临床误诊率。