Massively parallel sequencing of 10 autosomal STRs in Chinese using the ion torrent personal genome machine (PGM).
Massively parallel sequencing of 10 autosomal STRs in Chinese using the ion torrent personal genome machine (PGM).
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DOI：
10.1016/j.fsigen.2016.07.014
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发表时间：
2016-11
期刊：
影响因子：
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通讯作者：
Xueying Zhao;Hui Li;Z. Wang;K. Ma;Yu Cao;Wenbin Liu
中科院分区：
文献类型：
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作者：
Xueying Zhao;Hui Li;Z. Wang;K. Ma;Yu Cao;Wenbin Liu
大规模并行测序(MPS)技术越来越受到法医界的关注。MPS的高通量和同时分析多个标记的能力使其成为人类取证的一种有吸引力的方法。最近的研究表明，MPS系统在短串联重复序列(STR)分型中的应用是成功的。然而，不仅需要STR重复区域的DNA序列变异，而且还需要侧翼区域的DNA序列变异，以促进基于毛细管电泳分型方法的图谱共享。本研究通过设计168-273个碱基对的扩增片段，构建了MPS复合扩增体系，用于10个常染色体STR基因座(D13S317、D16S539、D19S433、D2S441、D3S1358、D5S818、D6S1043、D7S820、TH01、TPOX)的MPS分析。对165名中国无关个体的测序结果显示，扩增引物结合位点和核心重复序列之间的侧翼区有11个变异。此外，D13S317、D5S818和D7S820三个基因座在核心重复序列附近显示变异，导致基于序列和基于长度的分型结果不一致。4个基因座(D3S1358、D2S441、D19S433和D7S820)的MPS分型结果显示等位基因数目增加。本研究表明，MPS的STR分型可以提供更多来自STR基因座重复区和侧翼区的遗传信息，从而提高该系统在法医检测中的多样性和区分性。
Massively parallel sequencing (MPS) technology is gaining interest in the forensic community. The capabilities of high throughput and simultaneously analyses of many markers enable MPS as an attractive method for human forensics. Recent studies have demonstrated the successful application of the MPS system to short tandem repeat (STR) typing. However, not only DNA sequence variations in the repeat regions of STR but also in flanking regions are required to facilitate profiles sharing with capillary electrophoresis (CE)-based typing method. In this study, we constructed a multiplex PCR system for the MPS analysis of 10 autosomal STR loci (D13S317, D16S539, D19S433, D2S441, D3S1358, D5S818, D6S1043, D7S820, TH01, TPOX) by designing amplicons in the size range of 168–273 base pairs. Sequencing results from 165 Chinese unrelated individuals demonstrated 11 variations in the flanking regions between amplification primer binding sites and core repeat motifs. In addition, three loci, D13S317, D5S818, and D7S820, displayed variants adjacent to the core repeats and caused discordances between sequence-based and length-based typing results. Four loci (D3S1358, D2S441, D19S433 and D7S820) demonstrated an increased allele number using MPS-based typing. This study demonstrated that STR typing by MPS could provide more genetic information from both repeat and flanking regions of STR loci, thus improving the diversity and discrimination power of the system in forensic detection.