School of Forensic Medicine, Southern Medical University, Guangzhou, 510515, China.
AGCU ScienTech Incorporation, Wuxi, 214174, China.
Int J Legal Med. 2019 Jul;133(4):1015-1027. doi: 10.1007/s00414-018-1864-y. Epub 2018 May 30.
Y-chromosomal short tandem repeats (Y-STRs) have proven to be very useful in investigating sexual assault cases and in paternity lineage differentiation. However, currently available commercial Y-STR multiplex amplification systems bear the limitations in the identification of related males from the same paternal lineage due to there being an insufficient number of loci in any single amplification kit. The aim of this study was to establish and validate a novel 6-dye, 36-plex Y-STR multiplex amplification system that incorporated all of the loci present in the Yfiler™ Plus kit (DYS19, DYS385a/b, DYF387S1, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS449, DYS456, DYS458, DYS460, DYS481, DYS518, DYS533, DYS570, DYS576, DYS627, DYS635, Y_GATA_H4) as well as a further nine highly polymorphic Y-STR loci (DYS388, DYS444, DYS447, DYS522, DYS527a/b, DYS549, DYS596, DYS643). The novel system was optimized and validated by a series of studies that tested the effect of different PCR-based conditions as well as the species specificity, sensitivity, stability, stutter precision, suitability for use on DNA mixtures, reproducibility, and parallel testing of the system, as well as its performance on casework samples and population analysis, according to the SWGDAM developmental validation guidelines. A total of 246 haplotypes were found for the 36 Y-STRs among 247 Guangdong Han unrelated males. Collectively, the results demonstrate that the developed 36-plex Y-STR system is sensitive, robust, reliable, and highly informative for use in forensic genetics.
Y 染色体短串联重复序列（Y-STR）已被证明在性侵犯案件和亲子关系分化的研究中非常有用。然而，目前可用的商业 Y-STR 多重扩增系统由于在任何单一扩增试剂盒中存在的基因座数量不足，因此存在限制，无法识别来自同一父系的相关男性。本研究的目的是建立和验证一种新型的 6 色 36 plex Y-STR 多重扩增系统，该系统包含 Yfiler™Plus 试剂盒中存在的所有基因座（DYS19、DYS385a/b、DYF387S1、DYS389I/II、DYS390、DYS391、DYS392、DYS393、DYS437、DYS438、DYS439、DYS448、DYS449、DYS456、DYS458、DYS460、DYS481、DYS518、DYS533、DYS570、DYS576、DYS627、DYS635、Y_GATA_H4）以及另外 9 个高度多态性的 Y-STR 基因座（DYS388、DYS444、DYS447、DYS522、DYS527a/b、DYS549、DYS596、DYS643）。该新型系统通过一系列研究进行了优化和验证，这些研究测试了不同基于 PCR 的条件的影响，以及该系统的物种特异性、灵敏度、稳定性、突变精度、适用于 DNA 混合物、可重复性、以及该系统的平行测试，以及其在案例样本和群体分析中的表现，符合 SWGDAM 开发验证指南。在 247 名广东汉族无关男性中，36 个 Y-STR 共发现 246 个单倍型。总的来说，这些结果表明，开发的 36 plex Y-STR 系统在法医遗传学中具有敏感、稳健、可靠和高度信息丰富的特点。
