UNASSIGNED: ONT long-read sequencing was performed on patients with one reported PRKN/PINK1 pathogenic variant. EOPD patients with an age at onset younger than 50 were included in this study. As a positive control, we also included EOPD patients who had already been identified to carry two known PRKN pathogenic variants. Initial genetic testing was performed using either short-read targeted panel sequencing for single nucleotide variants and multiplex ligation-dependent probe amplification (MLPA) for copy number variants.
UNASSIGNED: 48 patients were included in this study (PRKN \"one-variant\" n = 24, PINK1 \"one-variant\" n = 12, PRKN \"two-variants\" n = 12). Using ONT long-read sequencing, we detected a second pathogenic variant in six PRKN \"one-variant\" patients (26%, 6/23) but none in the PINK1 \"one-variant\" patients (0%, 0/12). Long-read sequencing identified one case with a complex inversion, two instances of structural variant overlap, and three cases of duplication. In addition, in the positive control PRKN \"two-variants\" group, we were able to identify both pathogenic variants in PRKN in all the patients (100%, 12/12).
UNASSIGNED: This data highlights that ONT long-read sequencing is a powerful tool to identify a pathogenic structural variant at the PRKN locus that is often missed by conventional methods. Therefore, for cases where conventional methods fail to detect a second variant for EOPD, long-read sequencing should be considered as an alternative and complementary approach.
■对有1例PRKN/PINK1致病性变异的患者进行ONT长读测序。本研究包括发病年龄小于50岁的EOPD患者。作为阳性对照,我们还纳入了已被鉴定携带两种已知PRKN致病变种的EOPD患者.使用针对单核苷酸变体的短读取靶向组测序和针对拷贝数变体的多重连接依赖性探针扩增(MLPA)进行初始遗传测试。
■48例患者纳入本研究(PRKN\"单变异\"n=24,PINK1\"单变异\"n=12,PRKN\"双变异\"n=12)。使用ONT长读取测序,我们在6名PRKN“单变异”患者中检测到第二个致病变异(26%,6/23),但在PINK1“单变异”患者中没有(0%,0/12)。长读数测序确定了一个具有复杂反转的病例,结构变体重叠的两个实例,和三个重复的情况。此外,在阳性对照PRKN“双变”组中,我们能够在所有患者中鉴定出PRKN的两种致病变异(100%,12/12).
■该数据突出表明,ONT长读测序是一种强大的工具,可用于识别PRKN基因座上的致病性结构变异,而常规方法通常会错过这种变异。因此,对于常规方法无法检测到EOPD的第二种变体的情况,长读数测序应被视为一种替代和补充方法.