Xavier Catarina, Parson Walther
Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria.
Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria; Forensic Science Program, The Pennsylvania State University, PA, USA.
Forensic Sci Int Genet. 2017 May;28:188-194. doi: 10.1016/j.fsigen.2017.02.018. Epub 2017 Mar 1.
Massively Parallel (Next Generation) Sequencing (MPS) technologies have recently been proven useful and successful in typing various markers relevant in forensic genetics, such as STRs, SNPs and mitochondrial genomes. Early studies investigated self-developed DNA libraries, commercially supplied kits are currently being made available to allow a smoother and gradual implementation of such technologies in forensic laboratories. The ForenSeq™ DNA Signature Prep Kit (Illumina, CA) is the first commercially available STR kit that can be used on the MiSeq FGx™ (Illumina, CA) benchtop high-throughput sequencer. This kit allows the simultaneous typing of 59 STRs and up to 172 SNPs in a single reaction and presents a short library preparation protocol adapted to contemporary forensic requirements. In this study, we evaluated the beta version of the ForenSeq DNA Signature Prep Kit MiSeq FGx system by investigating reproducibility, sensitivity, mixtures, concordance, casework-type and aDNA samples and found it to perform successfully, proving to be a robust method for future forensic applications. MPS brings the possibility of complex multiplexing, high sensitivity and sequencing resolution to forensics; however, the need for consensual directions on databasing, data storage and nomenclature must be taken into consideration.
大规模平行（下一代）测序（MPS）技术最近已被证明在法医遗传学中对各类相关标记物进行分型时非常有用且成功，比如短串联重复序列（STR）、单核苷酸多态性（SNP）和线粒体基因组。早期研究调查的是自行开发的DNA文库，目前市面上有商业提供的试剂盒，以便法医实验室能更顺利且逐步地应用此类技术。ForenSeq™ DNA Signature Prep试剂盒（加利福尼亚州Illumina公司）是首个可用于MiSeq FGx™（加利福尼亚州Illumina公司）台式高通量测序仪的市售STR试剂盒。该试剂盒能在单个反应中同时对59个STR和多达172个SNP进行分型，并提供了一个适用于当代法医要求的简短文库制备方案。在本研究中，我们通过调查可重复性、灵敏度、混合样本、一致性、实际案例样本类型和古DNA样本，对ForenSeq DNA Signature Prep试剂盒MiSeq FGx系统的beta版本进行了评估，发现其表现成功，证明是未来法医应用的一种可靠方法。MPS为法医鉴定带来了复杂多重分析、高灵敏度和测序分辨率的可能性；然而，必须考虑在数据库建设、数据存储和命名法方面达成共识的指导方针的必要性。
