Y染色体单倍群SNP复合检测体系的构建及其在家系排查中的应用研究
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国家自然科学基金(81772027);国家重点研发计划(2017YFC0803501);公安部物证鉴定中心基本科研业务费专项资金项目(2017JB025);国家科技资源共享服务平台计划项目(YCZYPT[2017]01?3)


The development of Y chromosome haplogroup SNP multiplex system and application in familial searching
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    摘要:

    目的:构建Y染色体单倍群SNP复合检测体系(以下简称为“Y-SNP体系”),并评价其在法医遗传学家系排查中的应用价值。方法:筛选在东亚地区典型分布的Y染色体单倍群SNP位点,基于微测序技术构建复合体系并进行性能验证。采集4个社会学家系共20例男性血卡样本,利用Y-SNP体系与DNA TyperTM Y29试剂盒进行检测,统计分析同一家系中Y-SNP单倍群与Y-STR单倍型的差异,并利用网络图对其进行可视化。结果:筛选出74个Y-SNP位点并构建两组复合检测体系。Y-SNP体系分型准确,最小DNA检出量为0.14 ng,可用于检测血卡、口腔拭子、指甲、毛发(带毛囊)等常见检材。在非人源DNA样本中,除黑猩猩与恒河猴外,其余均未检出特异性峰型。P1家系Y-STR单倍型与Y-SNP单倍群均相同;P2家系Y-STR单倍型不同(容差≤5),Y-SNP单倍群相同;P3家系Y-STR单倍型不同(容差>5),Y-SNP单倍群不同;P4家系Y-STR单倍型不同(容差>5),Y-SNP单倍群相同。结论:Y-SNP体系与Y-STR数据联合应用可提高家系排查准确率。

    Abstract:

    Objective:This study aims to construct a Y chromosome haplogroup single nucleotide Polymorphism multiplex system(Y-SNP system)and evaluate its application value in familial searching of forensic genetic. Methods:The Y-SNPs that widely distributed in East Asia were screened. The system was constructed based on capillary electrophoresis(CE)genotyping platform,and its performance and accuracy were evaluated and verified. Twenty male samples from four pedigrees were collected. And the genotypes of Y-SNP and Y chromosome short tandem repeat(Y-STR)genetic markers were performed using the Y-SNP system and DNA TyperTM Y29 kit respectively. The differences of haplogroups and haplotypes in each pedigree were analyzed,and the network was used to visualize the mutation. Results:A total of 74 Y-SNPs were selected and constructed in double-tube of multiplex system. The SNP genotypes obtained by Y-SNP system were completely consistent with Sanger sequencing results. The low DNA detection limit was 0.14 ng. In addition,this system was suitable for blood stain,oral swabs,nails,hair root. Except chimpanzees and rhesus monkeys,there was no specific peak in non-human DNA samples. In P1 pedigree,the Y-SNP haplogroups and Y-STR haplotypes of samples were identical. In P2 pedigree,the Y-SNP haplogroups of all samples were identical and Y-STR haplotypes were different(mismatched loci ≤5). In P3 pedigree,the Y-SNP haplogroups of all samples were different and Y-STR haplotypes were different(mismatched loci >5). In P4 pedigree,the Y-SNP haplogroups of all samples were identical and Y-STR haplotypes were different(mismatched loci >5). Conclusion:The combination of Y-SNP and Y-STR can improve the accuracy of familial searching.

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宋正阳,王小娟,钱恩芳,师 源,董 薇,陈 飞,李彩霞,黄 江,江 丽. Y染色体单倍群SNP复合检测体系的构建及其在家系排查中的应用研究[J].南京医科大学学报(自然科学版),2021,(12):1759-1766

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  • 收稿日期:2021-09-30
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  • 在线发布日期: 2021-12-30
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