对于人类mtDNA的全频谱变异的HiFi长读幅子测序
Yan Lin1, Jiayin Wang1, Ran Xu2
1Research Institute of Neuromuscular and Neurodegenerative Diseases and Department of Neurology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, 250012, China.
BMC genomics
|May 31, 2024
概括
长读测序 (LRS) 与下一代测序 (NGS) 相比,可以更好地检测线粒体DNA (mtDNA) 变异,包括低百分比单核酸变异 (SNV) 和结构变异 (SV),相比下一代测序 (NGS). 这一进步改善了线粒体疾病 (MDs) 的诊断.
科学领域:
- 遗传学和基因组学 在
- 线粒体生物学 线粒体生物学
- 分子诊断学 分子诊断学
背景情况:
- 线粒体疾病 (MDs) 是由线粒体DNA (mtDNA) 的变异引起的.
- 下一代测序 (NGS) 在检测mtDNA中低百分比变异和删除方面面临限制.
- 精确检测线粒体基因组变异对于诊断MDs至关重要.
研究的目的:
- 评估长读测序 (LRS) 结合针对性远程聚合酶链反应 (LR-PCR) 的有效性,用于mtDNA变体检测.
- 为了比较LRS与传统NGS在mtDNA中识别单核酸变异 (SNV) 和结构变异 (SV) 的性能.
- 评估LRS在检测低频和二次mtDNA删除方面的能力.
主要方法:
- 对34名参与者 (28名患者,6名对照人) 的分析,使用向的LR-PCR和PacBio HiFi测序.
- 在17个样本中,LRS和NGS之间的变种检测的比较.
- 用桑格测序和肌肉纤维实时PCR量化验证LRS识别的删除.
主要成果:
- 在20名患者中,LRS检测到单个或多个删除变异 (比例>4%),并在2名患者中确定了m.3243 A>G热点变异.
- 在检测SNV频率低于5%的情况下,LRS表现优于NGS,并识别了更多的删除变异 (13/17个案例与8/17个案例).
- 在异常发炎性肌肉病 (IIM) 中,LRS准确地确定了二次mtDNA删除.
结论:
- 长读测序 (LRS) 在检测mtDNA中的各种SNV和SV,包括低频变异方面明显优于NGS.
- LRS为线粒体疾病提供了增强的诊断能力,为遗传变异提供了深刻的见解.
- 这项研究在了解mtDNA变异检测及其临床影响方面取得了重大进展.
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