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在DNA纳米球测序平台上使用长片段放大策略构建的mtDNA全基因组测序系统的法医效率评估

Man Chen¹, Chong Chen², Ning Li³

¹Guangzhou Key Laboratory of Forensic Multi‑Omics for Precision Identification, School of Forensic Medicine, Southern Medical University, Guangzhou 510515, China.

Forensic science international. Genetics

|August 31, 2024

概括

这项研究引入了一种用于法医识别的全线粒体DNA (mtDNA) 基因组测序系统. 该系统在分析包括降解样本在内的各种样本的mtDNA方面表现出高的准确性和效率.

科学领域:

遗传学遗传学是一个
法医科学法医科学法医科学
分子生物学分子生物学

背景情况:

线粒体DNA (mtDNA) 对于法医识别和人口研究至关重要,因为它的复制数量很高,母系遗传.
现有的mtDNA测序方法缺乏定量评估,导致解释不一致性和潜在错误,特别是与退化或混合DNA样本.
核基因组DNA (ngDNA) 污染和mtDNA序列/结构异质性对准确的mtDNA分析构成挑战.

研究的目的:

在DNA纳米球 (DNB) 测序平台上使用长片段放大策略评估一种全新的mtDNA基因组测序系统.
以测序质量,特异性,准确性和效率来评估系统的性能,在各种样本类型中用于法医应用.

主要方法:

在DNB测序平台上使用长片段放大策略进行全mtDNA基因组测序.
使用Q20和Q30值评估测序质量,并与全基因组DNA (wgDNA),mtDNA和核基因组DNA (ngDNA) 引用进行映射读取.
评估了使用已知的mtDNA单元型的人工混合物对异质质体的鉴定效果,并分析了基调的错误率.
在各种样本类型上测试了系统的性能,包括对照DNA,FTA血迹,天然样本 (FTA卡片/过纸上的血液),退化样本和母系血统样本.

主要成果:

这种新系统表现出高的测序质量,平均Q20 (97.17-97.37%) 和Q30 (91.93-92.48%) 值.
对于mtDNA的高映射百分比 (99.97-99.40%) 和ngDNA的低映射 (0.03-0.60%) 表示高特异性.
实现了精确的异质体检测,绝对误差在更高的异质体率下显著减少,并且稳定在500×测序深度以上.
该系统在各种样本类型中显示出一致的效率,包括老化的血迹和退化的DNA,证实了其强度.

结论:

评估的全mtDNA基因组测序系统在分析mtDNA序列信息方面表现出高性能.
该系统显示出对法医识别应用的巨大潜力,包括对像退化DNA这样具有挑战性的样品进行分析.
它的准确性和效率也支持其在母亲遗传研究和人口遗传结构研究中的实用性.

关键词:

在DNA纳米球测序平台上.降解样本的降解样本解释异质体的解释线粒体DNA全基因组测序

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