Villaescusa Patricia, Palencia-Madrid Leire, Campaner Magdalena Antònia, Jauregui-Rada Jaione, Guerra-Rodríguez Miguel, Rocandio Ana María, de Pancorbo Marian M
BIOMICs Research Group, Lascaray Research Center, University of the Basque Country UPV/EHU, Avda. Miguel de Unamuno, 3, 01006, Vitoria-Gasteiz, Spain.
Int J Legal Med. 2019 Jan;133(1):17-23. doi: 10.1007/s00414-018-1936-z. Epub 2018 Sep 18.
Single-nucleotide polymorphisms (SNPs) found within the non-recombining region of the Y chromosome (NRY) represent a powerful tool in forensic genetics for inferring the paternal ancestry of a vestige and complement the determination of biogeographical origin in combination with other markers like AIMs. In the present study, we introduce a panel of 15 Y-SNPs for a fine-resolution subtyping of the haplogroup R1b-DF27, in a single minisequencing reaction. This is the first minisequencing panel that allows a fine subtyping of R1b-DF27, which displays high frequencies in Iberian and Iberian-influenced populations. This panel includes subhaplogroups of DF27 that display moderate geographical differentiation, of interest to link a sample with a specific location of the Iberian Peninsula or with Iberian ancestry. Conversely, part of the intricacy of a new minisequencing panel is to have all the included variants available to test the effectiveness of the analysis method. We have overcome the absence of the least common variants through site-directed mutagenesis. Overall, the results show that our panel is a robust and effective method for subtyping R1b-DF27 lineages from a minimal amount of DNA, and its high resolution enables to improve male lineage discrimination in Iberian and Southwest European descent individuals. The small length of the amplicons and its reproducibility makes this assay suitable for forensic and population genetics purposes.
在Y染色体非重组区域（NRY）中发现的单核苷酸多态性（SNP）是法医遗传学中推断父系血统的有力工具，与其他标记（如AIMs）相结合可补充生物地理起源的确定。在本研究中，我们在单个微测序反应中引入了一组15个Y-SNP，用于对单倍群R1b-DF27进行高分辨率亚型分析。这是第一个能够对R1b-DF27进行精细亚型分析的微测序组，R1b-DF27在伊比利亚及受伊比利亚影响的人群中具有较高频率。该组包括DF27的亚单倍群，它们表现出适度的地理分化，有助于将样本与伊比利亚半岛的特定地点或伊比利亚血统联系起来。相反，新的微测序组的部分复杂性在于要拥有所有包含的变体以测试分析方法的有效性。我们通过定点诱变克服了最不常见变体的缺失问题。总体而言，结果表明我们的微测序组是一种从极少量DNA中对R1b-DF27谱系进行亚型分析的强大且有效的方法，其高分辨率能够提高对伊比利亚和西南欧血统个体的父系谱系区分能力。扩增子的短长度及其可重复性使得该检测方法适用于法医和群体遗传学目的。
