Multiplex Y-STRs analysis using the ion torrent personal genome machine (PGM).
Multiplex Y-STRs analysis using the ion torrent personal genome machine (PGM).
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DOI：
10.1016/j.fsigen.2015.06.012
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发表时间：
2015-11
期刊：
影响因子：
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通讯作者：
Xueying Zhao;K. Ma;Hui Li;Yu Cao;Wenbin Liu;Huaigu Zhou;Y. Ping
中科院分区：
文献类型：
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作者：
Xueying Zhao;K. Ma;Hui Li;Yu Cao;Wenbin Liu;Huaigu Zhou;Y. Ping
大规模平行测序（MPS）技术允许以期望的覆盖深度对多个样品的许多靶向区域进行平行测序分析。MPS在法医遗传学中的常规使用即将到来。本研究探讨MPS技术在法医学Y-STR分析中的应用。本研究设计了一种13个Y-STR基因座（DYS 19、DYS 389 I、DYS 389 II、DYS 390、DYS 391、DYS 392、DYS 437、DYS 438、DYS 439、DYS 448、DYS 456、DYS 635、GATA-H4）的多重PCR检测方法。对42名无关男性个体进行了多重Y-STR扩增，并使用离子激流个人基因组机器（PGM）系统对扩增产物进行了同步测序。所有基因座均被成功检测到，除了DYS 389 II由于相对较长的扩增子大小而表现出1.8%的失败率。在DYS 389 II、DYS 390、DYS 437、DYS 448和DYS 635中分别观察到7、3、2、6和5个新的等位基因，其中DYS 438中观察到1个新的等位基因。DYS 390的一个等位基因与传统毛细管电泳（CE）的等位基因识别不一致，因为核心重复单元上游有4个碱基对缺失。本研究表明，Y-STR分型的MPS可以提供更多的遗传信息，持有高的区分力的承诺。
Massively parallel sequencing (MPS) technologies allow parallel sequencing analyses of many targeted regions of multiple samples at desirable depth of coverage. Routine use of MPS for forensic genetics is on the horizon. In this study, we explore the application of MPS technology in forensic Y-STR analysis. We designed a multiplex assay with 13 Y-STR loci (DYS19, DYS389 I, DYS389 II, DYS390, DYS391, DYS392, DYS437, DYS438, DYS439, DYS448, DYS456, DYS635, GATA-H4) for the purpose of MPS. The multiplex Y-STR assay was amplified in 42 unrelated male individuals and amplicons were sequenced simultaneously using the ion torrent personal genome machine (PGM) system. All loci were detected successfully, except for DYS389 II that exhibited a failure rate of 1.8% due to the relatively long amplicon sizes. We observed 7, 3, 2, 6 and 5 new alleles, respectively in DYS389 II, DYS390, DYS437, DYS448 and DYS635 due to the presence of sub-repeat composition differences, and a new allele in DYS438 because of nucleotide substitution. One allele of DYS390 was inconsistent with allele call from conventional capillary electrophoresis (CE) because of 4 bp deletions upstream of the core repeat unit. This study demonstrates that Y-STR typing by MPS can provide more genetic information, holding the promise for high discriminatory power.