Department of Forensic Medicine, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Korea.
Brain Korea 21 PLUS Project for Medical Science, Yonsei University, 50-1 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Korea.
Sci Rep. 2021 Feb 25;11(1):4701. doi: 10.1038/s41598-021-82814-z.
The introduction of massively parallel sequencing (MPS) in forensic investigation enables sequence-based large-scale multiplexing beyond size-based analysis using capillary electrophoresis (CE). For the practical application of MPS to forensic casework, many population studies have provided sequence data for autosomal short tandem repeats (STRs). However, SE33, a highly polymorphic STR marker, has little sequence-based data because of difficulties in analysis. In this study, 25 autosomal STRs were analyzed, including SE33, using an in-house MPS panel for 350 samples from four populations (African-American, Caucasian, Hispanic, and Korean). The barcoded MPS library was generated using a two-step PCR method and sequenced using a MiSeq System. As a result, 99.88% genotype concordance was obtained between length- and sequence-based analyses. In SE33, the most discordances (eight samples, 0.08%) were observed because of the 4 bp deletion between the CE and MPS primer binding sites. Compared with the length-based CE method, the number of alleles increased from 332 to 725 (2.18-fold) for 25 autosomal STRs in the sequence-based MPS method. Notably, additional 129 unique alleles, a 4.15-fold increase, were detected in SE33 by identifying sequence variations. This population data set provides sequence variations and sequence-based allele frequencies for 25 autosomal STRs.
大规模平行测序（MPS）在法医调查中的引入使基于序列的大规模多重分析成为可能，超越了基于毛细管电泳（CE）的大小分析。为了将 MPS 实际应用于法医案例工作，许多群体研究已经提供了用于常染色体短串联重复序列（STRs）的序列数据。然而，由于分析困难，SE33 这个高度多态性的 STR 标记几乎没有基于序列的数据。在这项研究中，使用内部 MPS 面板分析了包括 SE33 在内的 25 个常染色体 STR，该面板对来自四个群体（非裔美国人、白种人、西班牙裔和韩国人）的 350 个样本进行了分析。使用两步 PCR 方法生成带条码的 MPS 文库，并使用 MiSeq 系统进行测序。结果，长度和基于序列的分析之间获得了 99.88%的基因型一致性。在 SE33 中，由于 CE 和 MPS 引物结合位点之间的 4 bp 缺失，观察到最多的不匹配（8 个样本，0.08%）。与基于长度的 CE 方法相比，基于序列的 MPS 方法中 25 个常染色体 STR 的等位基因数量从 332 增加到 725（增加 2.18 倍）。值得注意的是，通过识别序列变异，在 SE33 中检测到了额外的 129 个独特等位基因，增加了 4.15 倍。该群体数据集提供了 25 个常染色体 STR 的序列变异和基于序列的等位基因频率。
