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使用独特的分子标识符和SiMSen-Seq方法对STR标记物的超敏感测序
Maja Sidstedt1, Arvid H Gynnå1, Kevin M Kiesler2
1National Forensic Centre, Swedish Police Authority, Linköping SE-581 94, Sweden.
Forensic science international. Genetics
|April 10, 2024
概括
独特的分子标识符 (UMIs) 显著减少了大规模并行测序 (MPS) 中的错误,用于法医短串重复 (STR) 分析. 这种基于UMI的方法增强了法医样本中低DNA量和复杂混合物的检测.
科学领域:
- 法医科学 法医科学 法医科学
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
背景情况:
- 大规模并行测序 (MPS) 对于法医短串重复 (STR) 分析至关重要.
- PCR和测序错误,包括口吃器件,限制了低DNA量检测,特别是在复杂的混合物中.
- 独特的分子标识符 (UMIs) 在减少各种科学领域的测序噪声方面是有效的.
研究的目的:
- 开发和评估一种新的基于UMI的方法,SiMSen-Seq,用于使用MPS进行法医STR分析.
- 为UMI-STR测序创建一个概念验证七个STR复合体和相关的生物信息管道.
- 评估机器学习 (ML) 模型对改善UMI等位基调调用法医应用中的实用性.
主要方法:
- 采用了SiMSen-Seq方法,采用基于PCR的UMI引入与发针设计,以最大限度地减少原料结合和优化PCR协议.
- 一个七个STR多重测试被开发用于MPS库准备.
- 整合了机器学习模型,以提高基于UMI的等位基鉴定的准确性.
主要成果:
- 七个STR复合体在1 ng的单源样本和低模板样本下降到62.5 pg的样本中实现了完全检测和一致的等位基因.
- 对于具有挑战性的混合物 (10pg-150pg,115%的小贡献),使用UMI和ML过器检测到99.2%的预期基因.
- UMI显著降低了人工制造物和口吃率,通常低于父母等位基因的5%.
结论:
- 基于UMI的STR测序,特别是使用SiMSen-Seq方法,显著改善了具有挑战性的法医样本的分析.
- 这种方法提高了低DNA数量和复杂混合物的检测灵敏度和准确性.
- 减少文物和口吃,为法医调查提供了更可靠的STR资料.
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