PDF(Pubmed)
Abstract：
BACKGROUND: Paroxysmal kinesigenic dyskinesia (PKD) is the most prevalent kind type of paroxysmal Dyskinesia, characterized by recurrent and transient episodes of involuntary movements. Most PKD cases were attributed to the proline-rich transmembrane protein 2 (PRRT2) gene, in which the c.649 region is a hotspot for known mutations. Even though some patients with PKD have been genetically diagnosed using whole-exome sequencing (WES) and Sanger sequencing, there are still cases of missed diagnoses due to the limitations of sequencing technology and analytic methods on throughput.
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.
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.
摘要：
背景：阵发性运动障碍（PKD）是最常见的阵发性运动障碍类型，以不自主运动的反复发作和短暂发作为特征。大多数PKD病例归因于富含脯氨酸的跨膜蛋白2（PRRT2）基因，其中c.649区域是已知突变的热点。尽管一些PKD患者已经使用全外显子组测序（WES）和Sanger测序进行了基因诊断，由于测序技术和分析方法对通量的限制，仍有漏诊的情况。
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。
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。
