法医学杂志

法医学杂志 ›› 2024, Vol. 40 ›› Issue (1): 50-58.DOI: 10.12116/j.issn.1004-5619.2023.530803

• 技术与应用 • 上一篇    下一篇

42个微单倍型复合检测体系的构建及法医学应用

云鹏1,2(), 陈安琪3, 陈丽琴1(), 李成涛2()   

  1. 1.内蒙古医科大学法医学教研室，内蒙古 呼和浩特 010030
    2.司法鉴定科学研究院 上海市法医学重点实验室 司法部司法鉴定重点实验室 上海市司法鉴定专业技术服务平台，上海 200063
    3.复旦大学上海医学院基础医学院，上海 200032
  • 收稿日期:2023-08-29 发布日期:2024-02-18 出版日期:2024-02-25
  • 通讯作者: 陈丽琴,李成涛
  • 作者简介:云鹏（1988—），男，硕士研究生，主要从事法医遗传学研究；E-mail：491354640@qq.com
  • 基金资助:
    国家自然科学基金资助项目(81930056);内蒙古自治区自然科学基金资助项目(2019MS08155)

Establishment and Application of a 42-plex Microhaplotype Assay in Forensic Genetics

Peng YUN1,2(), An-qi CHEN3, Li-qin CHEN1(), Cheng-tao LI2()   

  1. 1.Department of Forensic Medicine, Inner Mongolia Medical University, Hohhot 010030, China
    2.Shanghai Key Laboratory of Forensic Medicine, Key Laboratory of Forensic Science, Ministry of Justice, Shanghai Forensic Service Platform, Academy of Forensic Science, Shanghai 200063, China
    3.School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China
  • Received:2023-08-29 Online:2024-02-18 Published:2024-02-25
  • Contact: Li-qin CHEN, Cheng-tao LI

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摘要：

目的 基于二代测序平台（next-generation sequencing，NGS）对42个微单倍型（microhaplotype，MH）遗传标记进行复合检测体系构建与法医学验证，探讨该体系在法医物证学实践中的应用价值。 方法 从前期研究中筛选42个高多态性MH并构建复合检测体系（42-plex MHs），通过MiSeq FGxTM平台进行测序，对该检测体系的重复性、灵敏度、特异性、稳定性、混合样本解析能力进行验证。通过对102例中国江苏省溧阳市汉族无关个体进行群体遗传学调查，评估该体系的法医学应用价值。 结果 该复合检测体系（42-plex MHs）的测序结果重复性可达100%，灵敏度低至0.062 5 ng，可耐受靛蓝（≤2 500 ng/μL）、腐殖酸（≤9 ng/μL）、血红素（≤20 μmol）、尿素（≤200 ng/μL）的干扰，并可鉴定2人（1∶19）、3人（1∶1∶9）和4人（1∶1∶1∶9）的混合样本。基于102例个体数据，42-plex MHs的累积个体识别率为1-3.45×10-30、累积非父排除率为1-3.77×10-11，平均有效等位基因数值为2.899。 结论 本研究成功构建了42-plex MHs复合检测体系，该体系具有高度的重复性与灵敏度，良好的抗干扰物能力与混合物解析能力，所含MH遗传标记具有较高的多态性，兼顾族源推断同时在个体识别和亲子鉴定相关案件中具有良好应用价值。

关键词: 法医遗传学, 微单倍型, 二代测序, 混合样本, 个体识别

Abstract:

Objective To establish and forensically verify a 42 microhaplotypes （mircohaps, MHs） multiplex assay system based on next-generation sequencing （NGS）, and to explore the application value of this system in the practice of forensic genetics. Methods A total of 42 highly polymorphic MHs were selected from previous studies, and sequenced by the MiSeq FGxTM platform to verify the repeata-bility, sensitivity, specificity, stability, and mixture analysis ability of the detection system. Through population genetic investigation of 102 unrelated Chinese Han individuals in Liyang City, Jiangsu Province, China, the application value of this system in forensic genetics was evaluated. Results The sequencing repeatability of the 42-plex MHs assay was 100% and the sensitivity was as low as 0.062 5 ng. The system had the ability to withstand the interference of indigo （≤2 500 ng/μL）, humic acid （≤9 ng/μL）, hemoglobin（≤20 μmol）, and urea （≤200 ng/μL） and to detect mixtures of 2 people （1∶19）， 3 people （1∶1∶9） and 4 people （1∶1∶1∶9）. Based on 102 individual data, the combined power of discrimination and the combined power of exclusion were 1-3.45×10-30 and 1-3.77×10-11, respectively, and the average effect value of alleles was 2.899. Conclusion The 42-plex MHs assay was successfully established in this study and this system has high repeatability and sensitivity, good anti-jamming ability and mixture analysis ability. The 42 MHs are highly polymorphism and have good application value in individual identification and paternity testing.

Key words: forensic genetics, microhaplotype, next-generation sequencing, mixture, individual identification

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