Background: Mutations in the bone morphogenetic protein receptor type 2 gene (BMPR2) represent a major genetic cause of pulmonary arterial hypertension (PAH). Identification of BMPR2 mutations is crucial for the genetic diagnosis of PAH. MinION nanopore sequencer is a portable third-generation technology that enables long-read sequencing at a low-cost. This nanopore technology-based device has not been used previously for PAH diagnosis. This study aimed to determine the feasibility of using MinION nanopore sequencing for the genetic analysis of PAH patients, focused on BMPR2.

Methods: We developed a protocol for the custom bioinformatics pipeline analysis of long reads generated by long-PCR. To evaluate the potential of using MinION sequencing in PAH, we analyzed five samples, including those of two idiopathic PAH patients and a family of three members with one affected patient. Sanger sequencing analysis was performed to validate the variants.

Results: The median read length was around 3.4 kb and a good mean quality score of approximately 19 was obtained. The total number of reads generated was uniform among the cases and ranged from 2,268,263 to 3,126,719. The coverage was consistent across flow cells in which the average number of reads per base ranged from 80,375 to 135,603. We identified two polymorphic variants and three mutations in four out of five patients. Certain indel variant calling-related errors were observed, mostly outside coding sequences.

Conclusion: We have shown the ability of this portable nanopore sequencer to detect BMPR2 mutations in patients with PAH. The MinION nanopore sequencer is a promising tool for screening BMPR2 mutations, especially in small laboratories and research groups.

背景： 骨形态发生蛋白受体2型基因突变（BMPR2) 代表肺动脉高压 (PAH) 的主要遗传原因。鉴定BMPR2突变对于 PAH 的基因诊断至关重要。MinION 纳米孔测序仪是一种便携式第三代技术，能够以低成本实现长读长测序。这种基于纳米孔技术的设备以前从未用于 PAH 诊断。本研究旨在确定使用 MinION 纳米孔测序对 PAH 患者进行基因分析的可行性，重点是BMPR2.

方法：我们开发了一个协议，用于对长 PCR 生成的长读数进行自定义生物信息学管道分析。为了评估在 PAH 中使用 MinION 测序的潜力，我们分析了五个样本，包括两名特发性 PAH 患者和一个三人家庭和一名受影响患者的样本。进行桑格测序分析以验证变体。

结果：中位读取长度约为 3.4 kb，获得了约 19 的良好平均质量评分。生成的读取总数在案例中是一致的，范围从 2,268,263 到 3,126,719。整个流动池的覆盖率是一致的，其中每个碱基的平均读取数在 80,375 到 135,603 之间。我们在五分之四的患者中发现了两个多态性变异和三个突变。观察到某些与 indel 变体调用相关的错误，主要是在编码序列之外。

结论： 我们已经展示了这种便携式纳米孔测序仪的检测能力 BMPR2PAH 患者的基因突变。MinION 纳米孔测序仪是一种很有前途的筛选工具BMPR2 突变，尤其是在小型实验室和研究小组中。