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下一代测序方法检测低频突变:"如果你能抓住我"

Vijay Menon¹, Douglas E Brash²

¹Department of Therapeutic Radiology, Yale School of Medicine, New Haven, CT 06520-8040, USA.

Mutation research. Reviews in mutation research

|September 16, 2023

概括

检测罕见的DNA突变对于健康至关重要,但具有挑战性. 新的超敏感测序方法显著改善了超低频突变的检测,揭示了常见方法往往产生虚假结果.

科学领域:

基因组学和分子生物学
生物信息学和计算生物学
临床诊断和预后研究

背景情况:

突变,或DNA的变化,在组织中以不同的频率发生,由于它们的稀有性,对临床应用提出了挑战.
标准下一代测序 (NGS) 在检测超低频突变 (VAFs <0.5%) 中存在局限性,阻碍了准确的诊断和研究.
不一致的术语和方法使不同研究小组对突变频率的可靠表征变得复杂.

研究的目的:

定义术语并描述测量超低频突变的挑战.
审查和描述最近在测序方法的创新,旨在提高罕见DNA变异的检测.
突出这些先进的测序技术的实际应用和影响.

主要方法:

复习和分类先进的测序技术,包括单链,双链和超敏感的父链共识测序方法.
详细描述了诸如Safe-SeqS,SiMSen-Seq,o2n-Seq,SMM-Seq,DuplexSeq,PacBio HiFi等方法,以及其他方法.
讨论量化变异性等位基因频率 (VAF) 到每核酸10-5和突变频率 (MF) 到每核酸10-7的方法,甚至通过分析大型基因组区域来降低这种方法.

主要成果:

超敏感测序方法使VAF的量化下降到10-5和MF的量化下降到10-7每核酸.
一些方法可以通过分析广泛的非重复部位来量化MF低于10-9/核酸或<15个错误/单 haploid基因组.
超敏感技术表明,在没有这些先进的方法的情况下,以前在0.5-1%的VAF中观察到的许多突变都是虚假的.

结论:

最近的测序技术创新显著提高了检测和量化超低频突变的能力.
这些超敏感的方法对于准确的临床诊断,预后,毒理学和识别新的疾病原因至关重要.
为了区分克隆扩张和独立突变,需要清楚报告突变频率指标 (MF_minI与MF_maxI).

关键词:

双重测序是指双重的测序.低频基因突变是一种低频的突变.这是下一代测序.罕见的变种罕见的变种变异性等位基因的频率变异

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